Special issue review article

Journal of Neuromuscular Diseases

Vol. 0(6): 1–16
Newborn screening and rapid genomic © The Author(s) 2024
Article reuse guidelines:
diagnosis of neuromuscular diseases sagepub.com/journals-permissions
DOI: 10.1177/22143602241296286
journals.sagepub.com/home/jnd

Tamara Dangouloff1 , Helena Lang2, Noor Benmhammed1

and Laurent Servais1,2

Abstract
Background: In recent years, treatments have been approved for certain neuromuscular diseases. In some cases, early
pre-symptomatic treatment is necessary for optimal response, and thus newborn screening is critical.
Objective: To review the current status of newborn screening programs for neuromuscular diseases and early diagnosis
through genetic testing.
Methods: Following the PRISMA guidelines, a literature search was performed on PubMed for screening of neuromus-
cular diseases; the search was conducted on literature available as of 1 May 2024.
Results: Included were 77 articles on newborn screening for seven diseases: spinal muscular atrophy (19 studies),
Duchenne muscular dystrophy (15), Pompe disease (20), X-linked adrenoleukodystrophy (14), Krabbe disease (6), meta-
chromatic leukodystrophy (2), and myotonic dystrophy 1 (1). Ten articles on rapid genomic diagnosis were identified.
Conclusion: Since 2021, newborn screening programs for neuromuscular diseases have been established, notably in X-linked
adrenoleukodystrophy, spinal muscular atrophy, Pompe disease, and Duchenne Muscular Dystrophy. Even in diseases where
treatment is currently not life-changing, such as Krabbe disease, new newborn screening programs continue to be implemented,
especially in the USA. The use of genetic diagnostic tests does not yet appear to be widespread or at least not widely reported. As
new treatments become available, genomic newborn screening programs will need to be rapidly and broadly implemented.

Keywords
neonatal screening, neuromuscular diseases, muscular atrophy, spinal, muscular dystrophy, duchenne, adrenoleukodystrophy,
early diagnosis, pompe disease, metachromatic leukodystrophy, myotonic dystrophy 1, krabbe disease
Received: 29 August 2024; accepted: 8 October 2024

Introduction the diagnostic odyssey.9 Diagnostic delays remain a real

issue, however, and are especially harmful for patients who
The treatment of patients with neuromuscular diseases (NMD)
suffer from treatable conditions. Newborn screening (NBS)
has evolved considerably in recent years as disease-modifying is a very efficient way of identifying treatable diseases. NBS
treatments have been approved for clinical use for patients has been in use for nearly 60 years for various diseases.
with several of the approximately 400 described NMDs. The
Incorporation of a test within an NBS program is based on a
diagnosis of an NMD is frequently delayed due to the rarity set of criteria proposed by Wilson and Jungner,10 with interpre-
and complexity of these diseases.1–4 This delay can result in tations varying from country to country, particularly as regards
irreversible muscle damage that can greatly reduce the effect-
iveness of treatment when available.5–7 Even in cases where
there is no muscle destruction, such as certain forms of con- 1
Neuromuscular Reference Center, Department of Pediatrics, University
genital myasthenia, the lengthy diagnostic process can lead Hospital Liege & University of Liege, Liege, Belgium
to decades of compromised quality of life before a correct 2
MDUK Oxford Neuromuscular Centre & NIHR Oxford Biomedical
diagnosis is made and suitable treatment is initiated as well Research Centre, University of Oxford, Oxford, UK
as secondary complications like scoliosis or chest infection.8
Corresponding author:
Improvements of the standard of care and diagnosis work- Tamara Dangouloff, CRMN, Hopital la Citadelle, Boulevard du XII de ligne,
flow, establishment of neuromuscular centers, and increased 4000 Liege, Belgium.
availability of genomic methods for testing have shortened Email: [email protected]

Creative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative Commons Attribution-
NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and
distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.
sagepub.com/en-us/nam/open-access-at-sage).
2 Journal of Neuromuscular Diseases 0(6)

to the criterion that a treatment be available. In Europe, for early diagnosis in symptomatic patients. Articles were
example, the approach is conservative with only diseases then sorted by disease.
that are treatable included, whereas in the United States of
America (USA) a more flexible approach has been adopted.11
NBS panels previously included only biochemical or meta- Results
bolic analyses and therefore could not include tests for diseases
without metabolic or endocrine markers such as NMDs.
Study selection process
Driven by the availability of effective treatment for certain The initial searches identified 772 articles describing NBS for
NMDs, genetic NBS is developing very rapidly. A notable neuromuscular disorders. They were screened by title and
example is spinal muscular atrophy (SMA). In just a few abstract, and 183 articles were identified for full-text screening.
years use of a genetic NBS test has become common in high- Full-text screening identified 105 articles; 21 articles were
income countries. The aim of this scoping review is to describe added based on reviews of bibliographies or after additional
the current status of NBS and early diagnosis of NMD. We searches of more recent publications. Of these 39 articles
carried out a scoping review over the last 10 years to assess were excluded because they described the same studies. In
the contribution of genetic testing by NBS or whole genome total 77 articles on screening for NMD and 10 on early diagno-
sequencing (WGS) for NMD. sis were included in this review. Supplementary file 2 shows the
flow chart based on the Preferred Reporting Items for
Methods Systematic Reviews and Meta-analyses (PRISMA) guidelines
used for the identification of these studies.
Literature search
We followed the PRISMA checklist12 and conducted a thorough Newborn screening programs for NMDs
literature search using Medline (PubMed). We sought to identify
Spinal muscular atrophy. SMA is a recessive disease caused
original, full-text articles that discussed NBS tests for NMDs that
by homozygous loss-of-function mutations (usually dele-
were published after 1 January 2014 and before 1 May 2024.
tions of exon 7) in SMN1, the gene encoding a protein crit-
To identify these articles, we used key terms related to
ical for maintenance of motor neurons. Three drugs are
NBS such as ‘neonatal screening’, ‘dried blood’, and
approved for treatment of SMA: nusinersen,13 onasemno-
‘Guthrie’, and those related to early diagnosis such as
gene-abeparvovec,14,15 and risdiplam.16 Extensive data
‘early diagnosis’, ‘whole genome sequencing’ in com-
demonstrated that these drugs are most efficacious if treat-
bination with relevant neuromuscular disease keywords.
ment begins before symptom onset.6,17
A detailed search strategy is provided in Supplementary
NBS for SMA was reported in 19 articles, which
file 1.
describe 19 studies each conducted in a region or
country.18–36 The initial implementation of pilot NBS pro-
Selection of studies grams for SMA took place in Taiwan in 2014,37 followed
by New York in 2016.38 Interestingly, these pilot programs
We used the Covidence application to screen the articles. To be were conducted before any drugs were approved for use in
included, articles had to be full articles, in English or French, these patients, and some of the patients identified in these
published after 1 January 2014 that had been peer reviewed. pilot programs were included in clinical trials.36,39 The inci-
Articles had to describe established NBS programs or pilot pro- dence rates vary significantly: For example, in Italy, a rate
jects for NMD or describe diagnoses made within the first 28 of 1 in 6000 was reported,40 whereas in Ontario the reported
days of a child’s life. Three reviewers (TD, HL, NB) first rate was 1 in 28,000.41 One of the lowest incidence rates of
screened titles and abstracts for eligibility with the consensus the disease was found in New York,42 which may be due to
of a minimum of two reviewers required for inclusion. Then increased use of preconception screening due to increased
two reviewers (TD, HL) reviewed the full text. Reasons for communication about the disease.43 In most programs, the
exclusions were recorded. Any potential disagreements were first tier of screening is done by quantitative real-time poly-
resolved by consensus. When more than one article described merase chain reaction (qPCR), and the second and con-
the same pilot project, only the most recent was included. We firmatory tests are by multiplex ligation-dependent probe
retained studies demonstrating the efficacy of NBS on deidenti- amplification (MLPA) or digital droplet PCR. Some pro-
fied Guthrie cards only if there were no pilot projects with iden- grams had issues with false positives, especially at the
tified patients. beginning of the program, and the reasons have been iden-
tified and corrected. For example, heparin was used in the tubes
used for sampling.28 In recent years, there has been exponential
Data extraction and presentation growth in the use of NBS for SMA, and most high-income
Data were extracted using the Covidence application. countries now screen newborns for this disease.44 A recent
Articles were then classified into two categories: NBS or survey showed that 100% of newborns in the USA and 66%
Dangouloff et al. 3

of newborns in Europe now screened for SMA.45 Table 1 sum- neurologically debilitating brain adrenoleukodystrophy
marizes the articles that describe NBS for SMA. enhances survival rates and improves functional outcomes.62
NBS for X-ALD was reported in 14 articles that describe
13 studies, each conducted in a different region or
Duchenne muscular dystrophy. Duchenne muscular dys-
country,63–67 most of them in the USA.62,68–75 Most of
trophy (DMD) is the most prevalent childhood form of mus-
these articles were published within the last five years, a
cular dystrophy. This condition arises from deletions,
sign of the rapid development of NBS programs of
duplications, or single nucleotide variants that affect the pro-
X-ALD. In the USA, the program, which was first imple-
duction of functional dystrophin. First symptoms typically
mented in 2016, has proven to be effective. The first-tier
occur when children are 2–3 years old and include proximal
tests for X-ALD are all metabolic, but confirmatory tests
weakness and muscle pseudo-hypertrophy. The weakness pro-
are increasingly genetic. The incidence of X-ALD in
gresses rapidly and results in the loss of walking between the
males varies from 1 in 400076 to 1 in 14,000.69 Table 3 sum-
ages of 7 and 15. On average, there is a two-year gap between
marizes the articles that describe NBS for X-ALD.
the onset of symptoms and diagnosis of DMD.1 This diagnos-
tic delay not only causes distress for parents, but it can also
Pompe disease. Pompe disease, also called glycogenosis type
result in a delay in appropriate care like steroids and multidis-
2, is an autosomal recessive lysosomal storage disorder result-
ciplinary follow-up.
ing from acid alpha-glucosidase (GAA) deficiency. The
The Food and Drug Administration (FDA) has approved
disease has a broad clinical spectrum, ranging from the infant-
eight drugs for use in patients with DMD: two steroids (deflaza-
ile form, which manifests within the first few months of life, to
cort and vamorolone), four oligo antisense nucleotide that aim to
adult forms. Without treatment, the infantile form invariably
skip exons 51 (eteplirsen, vitolarsen), 53 (golodirsen) and 45
results in early mortality due to cardiorespiratory failure or
(casimersen), a gene therapy that provides a copy of microdystro-
respiratory infection, typically before the age of one year. In
phin using an AAV-microdystrophin construct, and the histone
the later-onset forms, symptoms can emerge at any age and
deacetylase inhibitor givinostat. There are age restrictions on
are linked to progressive skeletal muscle dysfunction.
some of these drugs. For example, givinostat is for use in patients
Enzyme replacement therapy (ERT) with alglucosidase
6 years and older and the gene therapy is for use in patients 4
alfa (Myozyme) received approval from the FDA and the
years and older. The exon skipping drugs do not have age restric-
European Medicines Agency (EMA) in 2006.78 In 2023,
tions. Clinical and pre-clinical efforts toward development of a
the FDA approved the combination therapy of cipaglucosi-
number of additional potential therapies are ongoing.46
dase alfa-atga with miglustat to treat adults who are not
Our literature search identified 13 pilot projects or official
improving on their current regime of ERT.79 Early interven-
implementations of NBS for DMD between 1974 and 2022
tion with ERT is associated with improved outcomes.80
described in 15 articles.47–60 New pilot programs were recently
Gene therapy development is mostly focused on restoring
launched in the USA; the main objectives are to evaluate the
GAA production and is currently in clinical development.
feasibility and benefit of NBS for DMD.51,61 In the USA,
NBS for Pompe disease was reported in 20 articles
there is considerable pressure from parent associations to add
describing 18 studies. The inaugural NBS pilot program
DMD to the list of diseases in the Recommended Uniform
was launched in Taiwan in 2005 and has been consistently
Screening Panel (RUSP), even though evidence for efficacies
documented since.81,82 Since 2015, Pompe disease has been
of available treatments is limited. One of the rational of this pos-
included in the RUSP in the USA and screening for Pompe
ition is to avoid recurrence of the disease in the family. In
disease is part of NBS programs in 38 states.83 These efforts
screening efforts reported to date, the incidence in young
have been described in a number of publications.84–92
males is between 1 in 350048 to 1 in 10,300.49 Table 2 sum-
Comparable initiatives have been established in Italy,93
marizes the articles that describe NBS for DMD.
Mexico,94 Japan,95,96 China,97 and Brazil.98–100 The
reported incidences for infantile and late forms vary from
X-linked adrenoleukodystrophy. X-linked adrenoleukodystro- 1:10,600 in Brazil to 1:38,000 in Japan. Table 4 summarizes
phy (X-ALD) is a peroxisomal impairment caused by a loss the articles that describe NBS for Pompe disease.
of function mutation in the ABCD1 gene located on the X
chromosome. This metabolic disorder affects the adrenal Krabbe disease. Krabbe disease is an autosomal recessive
glands, brain, and spinal cord, resulting in devastating con- lysosomal disorder that impacts the white matter of both
sequences. Boys who are hemizygous are more severely the central and peripheral nervous systems. The severity
affected by X-ALD than are heterozygous girls (60% of depends on the age of onset. This disease is caused by
cases). If left untreated, subjects with X-ALD typically loss-of-function mutations in both alleles of the GALC
survive only a few years after symptom onset. Treatments gene, leading to a deficiency in galactosylceramidase.
include corticosteroids for adrenal insufficiency, hematopoi- Historically, 85–90% of patients are diagnosed with the infant-
etic stem cell transplantation, and gene therapy. Treatment ile form, the most severe type, which manifests within the first
during the initial stages of brain inflammation prior to six years of life. In cases where the disease begins within the
 4

Table 1. NBS for SMA.

Country (region) Date Pop Total number Scr First-tier Scr 2nd-tier Conf Cons First result N° cases Prev Aim of study
18
Taiwan 2014-2019 NB 364,000 qPCR ddPCR MLPA opt-in 21 21 1/17,000 Explore SMA NBS and treatment
of identified subjects
USA (NY)19 2016–2017 NB 3826 qPCR qPCR opt-in 1 1 1/3826 To determine feasibility of SMA NBS
Germany 20 2018–2020 NB 297,163 qPCR MLPA opt-in 43 43 1/6910 Report on 2-year pilot of SMA NBS
Belgium (FWB) 36 2018–2021 NB 136,339 qPCR qPCR MLPA MLPA opt-out 9 9 1/13,364 Explain start of SMA NBS
Australia (NSW/ACT) 21 2018–2021 NB 252,081 qPCR ddPCR ddPCR opt-out 22 21 1/11,458 Evaluate implementation of SMA NBS
USA (NC) 22 2018–2020 NB 12,065 qPCR qPCR qPCR opt-in 2 1 1/12,065 Evaluate SMA NBS
Italy (Tuscany, Lazio) 23 2019–2021 NB 90,885 qPCR qPCR qPCR opt-in 15 15 1/6059 Evaluate SMA NBS for extension
USA (GA) 29 2019–2020 NB 301,418 qPCR N/A N/A opt-out 39 15 1/18,840 Results of SMA NBS program: incidence, timing
USA (WI) 31 2019–2020 NB 60,984 qPCR ddPCR ddPCR opt-out 6 6 1/10,164 Report on 1-year SMA NBS program
Canada (Ontario) 26 2020–2021 NB 139,810 Mass Array MLPA opt-out 5 5 1/27,960 Report on beginning of SMA NBS
Japan (Osaka) 24 2020–2023 NB 105,419 qPCR MLPA MLPA opt-in 1 1 Evaluate SMA NBS
USA (CA) 25 2020–2021 NB 628,791 qPCR ddPCR PCR opt-out 34 34 1/18,494 Report on beginning of SMA NBS
Japan (Hyogo) 28 2021–2023 NB 16,037 qPCR MLPA qPCR opt-in 17 3 1/5346 Report on 2.5-year program of SMA NBS
Japan (Kumua-moto) 30 2021–2022 NB 13,587 qPCR qPCR MLPA opt-in 1 1 1/13,587 Report on SMA NBS program
Russia (8 regions) 32 2022 NB 202,908 qPCR qPCR MLPA opt-in 32 26 1/7804 Report on SMA NBS program
Russia (St Petersburg) 33 2022 NB 36,140 Genome PCR opt-in 4 4 1/9035 Report on SMA NBS program
X real-time
PCR-based
Ukraine 27 2022–2023 NB 65,880 qPCR MLPA opt-in 11 11 1/5989 Report on implementation of SMA NBS program
Croatia 34 2023 NB 32,655 qPCR qPCR MLPA opt-out 5 5 1/6531 Report on SMA NBS program
Serbia 35 2023 NB 54,393 qPCR MLPA MLPA opt-out 2 2 1/9066 Report on implementation of SMA NBS program
and results of the national program.
All 2,759,988 268 222 1/12,432

List of abbreviations: Conf: Confirmatory assay; Cons: parents’ method of consent; ddPCR: digital droplet PCR; First result: first result positive or inconclusive; FWB: Federation Wallonia-Brussels (Region of
Belgium); CA: California; GA: Georgia; MLPA: multiplex ligation-dependent probe amplification; NB: Newborn; N° cases: Number of confirmed cases; N/A: not available; NC: North Carolina, NSW/ACT: New
South Wales and Australian Capital Territory (region of Australia); NY: New-York; Pop: description of the population screened; Prev: Prevalence; qPCR: quantitative real-time polymerase chain reaction; Ref:
references used; Scr: Screening assay; WI: Wisconsin.
Journal of Neuromuscular Diseases 0(6)
 Dangouloff et al.

Table 2. NBS for DMD.

Total Scr Scr First N°

Country Date Pop number First-tier 2nd-tier Conf Cons result cases Prev Aim of study
47,48
Germany 1974–2011 NB (M) 537,000 CK assay / Opt-in 155 1/3500 Evaluation of DMD NBS
France 53 1975–1986 NB (M) 218,851 CK assay / Clinical assessment N/A 48 1/4600 Report on DMD NBS program
New Zealand 52 1979 NB (M) 10,000 CK assay / Clinical assessment N/A 2 1/5000 Report on DMD NBS program
Canada (Manitoba) 1986–2007 NB (M) 172,860 CK assay / DMD molecular testing Opt-out 18 1/9600 Determine whether screening reduces
47,57
number of second children born with
DMD. Observation of development.
49
USA (PA) 1987–1995 NB (M) 403,576 CK assay CK DMD molecular testing / Opt-out, 39 1/10,300 Description of attitudes of parents and of
isozyme muscle biopsy verbal patients with diagnosis was with DMD
consent NBS or not.
UK (Wales) 50 1990–2011 NB (M) 369,780 CK assay / DMD molecular testing / Opt-in 145 56 1/6600 Report on 21-year DMD NBS pilot
plasma CK program.
56
Cyprus 1992–1997 NB (M) 30,014 CK assay / DMD molecular testing / Opt-out 43 5 1/6000 Evaluation of the methods and
muscle biopsy implementation of DMD NBS pilot
program.
55
USA (OH) 2007–2010 NB (M) 17,865 CK assay / DMD molecular testing Opt-in 168 3 1/6000 Evaluate of methods and feasibility of
DMD NBS.
54
China (Zhejiang) 2017 NB (M) 18,424 CK-MM / DMD molecular testing Opt-in 13 4 1/4600 Recommendations for follow-up care.
assay
59
China (Henan) 2019–2019 NB (M) 13,110 CK-MM MLPA NGS and sanger Opt-in 3 3 1/4370 Determine optimal cutoff value and
sequencing evaluate assess detection rate.
51
USA (NY) 2019–2021 NB 36,781 (M: CK-MM CK-MM NGS Opt-in 42 3 M: 1/6218 Evaluate the feasibility and benefit of
18,654) DMD NBS.
58
USA (NC) 2020–2022 NB 13,354 (M: CK-MM Total CK NGS Opt-in 84 2 M: 1/3380 Implement and evaluate DMD NBS.
6759)
Taiwan 60 2021–2021 NB 50,572 (M: CK-MM CK-MM WES Opt-out 632 3 M: 1/8710 Establish that DMD NBS enables earlier
26,130) management.
All M: 1,843,023 1392 341 M: 1/5404

List of abbreviations: CK-MM: creatine kinase muscle; First result: first result positive or inconclusive; Conf: Confirmatory assay; Cons: parents’ method of consent; First result: Positive first result or
inconclusive M: male; N/A: not available; NGS: next-generation sequencing; NY: New York; NB: newborn; N° cases: Number of confirmed cases; NGS: Next-generation sequencing; OH: Ohio; PA: Pennsylvania;
Pop: description of the population screened; Prev: Prevalence; Ref: references used; Scr: Screening assay; WES: whole exome sequencing.
5
 6

Table 3. NBS for X-ALD.

Coun-try Date Pop Total number Scr First-tier Scr 2nd-tier Conf Cons Result N° cases Prev Aim of study
75
USA (NY) 2014–2015 NB 365,000 FIA-MS-MS Sanger sequencing N/A 33 M: 13 F: 13 All: 1/28,076 Report advisory committee
(4 Zellweger recommendation.
Syndrome)
Taiwan 63 2016 NB 320,528 FIA-MS-MS HPLC-MS-MS WGS Opt-in 43 M: 12 M: 1/13,825 Evaluate extended national NBS program.
69
USA (CA) 2016–2020 NB 1,854,631 FIA-MS-MS LC-MS-MS Sanger sequencing Opt-out 355 M: 95 F: 110 M: 1/14,397 Describe NBS experience.
(23
Zellweger
Syndrome)
76
USA (MN) 2017–2018 NB 67,836 FIA-MS-MS / Molecular testing Opt-out 56 M: 9 F: 5 All: 1/4845 Report on NBS.
+ Very-Long Male: 1/
Chain Fatty 3878
Acids analysis
70
USA (GA) 2017 NB 51,081 FIA-MS-MS LC-MS-MS Gene sequencing Opt-out 11 (M: 4, F: 7) M: 1 F: 3 All: 1/51,081 Report on use of two-tiered strategy
of NBS for ALD
71
USA (PA) 2017–2021 NB 542,554 FIA-MS-MS LC-MS-MS NGS of the ABCD1 Opt-out 51 M: 21 F: 23 M: 1/13,110 Review data and outcomes of NBS
(M: gene program
278,330)
USA (NC) 2018 NB 52,301 HPLC-MS-MS Sanger sequencing Opt-out 12 3 1/10,000 Describe NBS pilot.
72,77

USA (NE) 73 2018–2021 NB 82,920 FIA-MS-MS HPLC-MS-MS Gene-specific Opt-out 23 (M: 9 F: M: 4 F: 9 M: 1/10,583 Summarize initial experience with NBS.
(M: 42,332) long-range PCR 14)
for genomic
sequencing
USA (IL) 74 2019–2021 NB 276,000 LC-MS-MS Measurement Gene sequencing Opt-out 18 (M: 7, F: M: 4 F: 8 1/16,200 Report outcomes of initial NBS program.
of plasma 11)
VLFCA
China 64 2020–2021 NB 43,653 LC-MS-MS NGS Opt-in 32 (M: 18F: M: 7 F: 7 1/6236 Evaluate performance NBS program.
14) (2 other
diseases)
Italy 65 2021–2024 NB started FIA-MS-MS UHPLC NGS Opt-in / / / Improve diagnosis and subsequent and
MS-MS treatment.
66
Japan 2021–2025 NB 1000 HPLC-MS-MS HPLC-MS-MS VLCFA and ABCD1 Opt-in 0 0 / Improve prognosis of patients.
(M) genetic analyses
The Nether- 2021 NB 71,208 FIA-MS-MS HPLC-MS-MS Gene sequencing Opt-in 507 (M: 249) 4 All: 1/18,000 Report on boys-only pilot study of NBS.
lands 67 M: 1/9000
All All: 3,728,712 M: 837 M: 169 All: 1/22,063

List of abbreviations: CA: California; Conf: confirmatory assay; Cons: parents’ method of consent; F: female; FIA-MS-MS: flow injection analysis tandem mass spectrometry; First result: first result positive or
inconclusive; GA: Georgia; HPLC: high-performance liquid chromatograph; IL: Illinois; LC-MS-MS: liquid chromatography coupled to tandem mass spectrometry; M: male; MN: Minnesota; MS-MS: tandem mass
spectrometry; N° cases: Number of confirmed cases; NB: newborn; NC: North Carolina; NE: Nebraska; NGS: next-generation sequencing; PA: Pennsylvania; Pop: description of the population screened; Prev:
prevalence; Ref: references used; Scr: screening assay; UHPLC: ultra-high performance liquid chromatography; VLFCA: very long chain fatty acid.
Journal of Neuromuscular Diseases 0(6)
Table 4. NBS for pompe disease.

Total First
Coun-try Date Pop number Scr First-tier Scr 2nd-tier Conf Cons result N° cases Prev Aim of study
81
Taiwan 2005– NB 994,975 GAA activity / lymphocyte GAA Opt-in 55 16 IOPD 39 IOPD: 1/62,186 LOPD: Demonstrate advantage of early
Dangouloff et al.

2018 activity / GAA WGS LOPD 1/25,512 All: 1/ diagnosis and treatment, even for
18,090 LOPD.
Italy 93 2015– NB 112,446 FIA-MS-MS LC-MS/MS Enz + Mol test + Urinary Opt-in 28 2 IOPD 6 IOPD: 1/56,223 LOPD: Report on NBS regional program.
2018 GAGs analysis + CK LOPD 1/18,741 All: 1/
14,055
Taiwan 82 2010– NB 1,228,539 GAA activity GAA gene sequencing Opt-in 33 33 IOPD IOPD: 1/37,228 Demonstrate diagnostic and
2021 treatment strategies for IOPD.
Mexico 94 2012– NB 20,018 GAA activity / Enz + Mol test Opt-in 19 1 All: 1/20,000 Evaluate the results of a lysosomal
2016 NBS.
84
USA (WA) N/A NB 110,000 GAA activity MS/ GAA gene sequencing N/A 13 4 All: 1/27,800 To assess the performance of MS/MS.
MS
85
USA (NYS) 2013– NB 19,197 LC-MS-MS Opt-in 6 0 IOPD 1 1/19,197 Report on pilot NBS program.
2014 LOPD
USA (MO) 101 2013– NB 467,000 Fluor Digital / Enz + Mol test + Urinary Opt-out 274 10 IOPD 36 IOPD: 1/46,700 LOPD: Report on 6-year NBS program.
2018 Micro-fluidics GAGs analysis + CK LOPD 1/13,000 All: 1/
10,200
95,96
Japan 2013– NB 296,759 Fluor assay / NGS Opt-in 154 1 IOPD 7 IOPD: 1/296,759 Summarize results of NBS program.
2020 LOPD LOPD: 1/42,394 All:
1/38,094
USA (IL) 102 2015– NB 684,290 LC-MS-MS 2-tiered cut-off Enz + Mol test + Urinary Opt-out 397 3 IOPD 26 IOPD: 1/228,100 Description of NBS program.
2019 system analysis LOPD LOPD: 1/26,300 All:
1/23,600
87
USA (PA) 2016– NB 531,139 FIA-MS-MS / Enz + Mol test Opt-out 115 2 IOPD 31 IOPD: 1/265,500 Evaluation of benefits and challenges
2019 LOPD LOPD: 1/17,100 All: of NBS.
1/16,100
Brazil (Porto 2016-N/ NB 10,527 DMF NGS Opt-out N/A O IOPD 1 All: 1/10,527 Evaluation of challenges of NBS.
Alegre) 98,99 A LOPD
USA (KY) 88 2016– NB 55,161 FIA-MS-MS CLIR tools Opt-out 15 2 LOPD: 1/27,581 All: 1/ Report NBS program.
2017 27,581
USA (WI) 92 2017– NB 99,018 N/A N/A N/A N/A 12 0 IOPD 12 All: 1/8251 Report on pilot NBS program.
2019 LOPD
USA (MN) 89 2017– NB 257,726 FIA-MS-MS FIA-MS-MS GAA sequencing Opt-out 21 2 IOPD 15 OIPD: 1/128,863 Evaluate diagnostic and follow-up
2021 LOPD LOPD: 1/17,182 All: outcomes.
1/15,160
90
USA (CA) 2018– NB 453,152 FIA-MS-MS Mol test Mol test Opt-out 88 2 IOPD 16 IOPD: 1/226,600 Report on 1-year NBS program.
2019 LOPD LOPD: 1/28,300 All:
1/25,200
100
Brazil 2018 –N/ NB 20,066 FIA- MS-MS NGS Opt-out 8 0 IOPD 5 All: 1/10,600 Evaluation of challenges of NBS.
A LOPD
91
USA (GA) N/A NB 59,332 FIA- MS-MS FIA- MS-MS Opt-out 5 1 IOPD 2 IOPD: 1/59,332 LOPD: Evaluate implementation of NBS.
LOPD 1/29,666 All: 1/
19,777
7

(continued)
8 Journal of Neuromuscular Diseases 0(6)

first year, rapid neurological decline is observed, often resulting

first result positive or inconclusive; GA: Georgia; GAA: acid α-glucosidase; IOPD: infantile-onset Pompe disease; LC-MS-MS: liquid chromatography coupled to tandem mass spectrometry; LOPD: late-onset
List of abbreviations: Conf: Confirmatory assay; Cons: parents’ method of consent; DMF: digital micro-fluidics; Enz: Enzymatic testing; FIA-MS-MS: flow injection analysis tandem mass spectrometry; First result:
Evaluate the birth prevalence of
in death before the age of two. Late-onset Krabbe disease pre-

Pompe disease; Mol Test: molecular testing; MS-MS: tandem mass spectrometry; N/A: not available; NB: newborn; NGS: next-generation sequencing; Pop: description of the population screened; Prev:
and determine subclinical
sents with much more variability in symptoms and progres-
sion.103 The rarity of the disease poses a challenge for
evaluating the effectiveness of pre-symptomatic treatments.
Krabbe disease has proven resistant to most single-therapy
treatments. Hematopoietic stem cell transplantation, the
forms. current standard of care, can modify the disease’s progression,
but it only slows it down, even if begun pre-symptomatically.
Prev

Recent successes with combination therapies in small animal

models and the discovery of new pathogenic mechanisms
LOPD: 1/27,347

offer hope for developing effective treatments for this devastat-

ALL: 1/19,123
LOPD: 1/16,702

IOPD: 1/75,964

ing condition.104,105
Six studies have evaluated NBS for Krabbe disease: three in
N° cases

the USA,88,106,107 one in Mexico,94 one in China,97 and one in

Brazil.100 Given the low incidence of the disease, estimated at 1
in 100,000,105 not all programs have identified cases. Attempts
LOPD: 200

to include Krabbe on the RUSP have been twice unsuccessful

First result

All: 286

and in January of 2024, the Advisory Committee on

IOPD: 72
3 LOPD

Heritable Disorders in Newborns and Children (ACHDNC)

officially recommended that Krabbe disease be added to the
RUSP. It is currently implemented in only eleven states in the
Cons

1254

USA. Recent publications have raised ethical concerns regard-

Opt-in 10

ing such screening,108 even though patient associations are

pushing for its implementation.109 Table 5 summarizes the arti-
Conf

cles that describe NBS for Krabbe disease.

Metachromatic leukodystrophy. Metachromatic leukodystro-

phy (MLD) is a metabolic disorder inherited in an autosomal
recessive manner, characterized by the accumulation of sulfa-
Scr First-tier Scr 2nd-tier

tides in the central and peripheral nervous systems due to a defi-

ciency in the enzyme arylsulfatase A, resulting in
NGS

demyelination. The disease primarily manifests as a decline

in motor or cognitive functions. Depending on the subtype,
Prevalence; Ref: references used; Scr: Screening assay; Urinary GAGs analysis.

the disease has a highly variable progression and duration

but eventually leads to severe disability and death. Both the
Pompe

EMA and the FDA have approved atidarsagene autotemcel,

an autologous hematopoietic stem cell gene therapy, for the
treatment of children diagnosed with pre-symptomatic late
Total number

FIA- MS-MS

infantile, pre-symptomatic early juvenile, or early symptom-

atic early juvenile MLD.
The first pilot project aimed at evaluating the feasibility
of NBS for MLD was conducted in the USA in 2020,
5,469,453

screening 27,335 de-identified dried blood spots.110 Very

NB 50,108

recently, a study in Manchester, UK showed the effective-

Pop

ness of a sulphate test on dried blood spots and the evalu-

ation of ARSA enzyme activity as a two-tier strategy.111
One infant was identified. This program could soon be recom-
mended by the UK National Screening Committee for incorp-
Table 4. Continued.

2021
Date

oration into routine NBS. Table 5 summarizes the articles that

describe NBS programs for MLD.
(Shangai)97
Aim of study
Coun-try

Myotonic dystrophy 1. Myotonic dystrophy 1 (MD1) is an

China

autosomal dominant disorder marked by muscle weakness,

All

myotonia, early onset cataracts, and various systemic

 Dangouloff et al.

Table 5. NBS for Krabbe disease, MLD, and MD1.

Total First
Disease Coun-try Date Pop number Scr First-tier Scr 2nd-tier Conf Cons result N° cases Prev Aim of study
106
Krabbe USA (NY) 2006– NB 2,090,910 LC-MS-MS Mol test Enz Opt-out 620 5 1/418,000 Report on NBS program.
2014
Mexico 94 2012– NB 20,018 LC-MS-MS / Enz and Mol Test Opt-in 38 0 Evaluation of results of lysosomal
2016 NBS.
88
USA (KY) 2016– NB 55,161 FIA-MS-MS CLIR tools Opt-out 181 1 1/55,000 Report on NBS program.
2017
USA (IL) 107 2017– NB 494,147 LC-MS-MS Mol test + Follow-up Opt-out 288 2 IOKD 6 IOKD: 1/ Report on NBS program and role
2020 psychosine LOKD 250,000 of psychosine in disease
levels LOKD: 1/ diagnosis.
82,400 All:
1/61,800
100
Brazil 2018-N/ NB 20,066 MS-MS UHPLC MS-MS NGS Opt-out 1 0 Evaluation of challenges of NBS.
A
China (Shangai) 2021 NB 50,108 FIA-MS-MS (GALC) Krabbe NGS Opt-in 20 9 LOKD LOKD: 1/5567 Evaluation of birth prevalence and
97
prevalence of subclinical forms.
MLD USA (WA) 110 DBS 27,335 LC-MS-MS Enz Mol test no 195 2 Assessment of feasibility of NBS.
UK 2023-N/ DBS 3687 Sulphatides enz test - PCR: ARSA gene Enz sulphamidase and no 11 1 1/3687 Assessment of feasibility of NBS.
(Manchester) A UPLC-MS-MS sequencing β-galactosidase
111
112
MD1 USA (NY) 2013– DBS 51,341 triplet primed-PCR + Mol test no 143 24 1/2100 Determination of prevalence.
2014 melt curve analysis

List of abbreviations: Conf: Confirmatory assay; Cons: parents’ method of consent; DBS: deidentified; Enz test: enzymatic testing; FIA-MS-MS: flow injection analysis tandem mass spectrometry; First result: first
result positive or inconclusive; IOKD: infantile-onset Krabbe disease; LC-MS-MS: liquid chromatography coupled to tandem mass spectrometry; LOKD: late-onset Krabbe disease; Mol Test: molecular testing;
MS-MS: tandem mass spectrometry; N° cases: Number of confirmed cases; N/A: not available; NB: newborn; NGS: next-generation sequencing; Pop: description of the population screened; Prev: Prevalence;
Ref: references used; Scr: Screening assay; UHPLC MS-MS: ultra-high-performance liquid chromatography MS-MS; UPLC MS-MS: ultra-performance liquid chromatography MS-MS.
9
 10
Table 6. Rapid genomic diagnosis of NMD.
Country Date Pop Total number Scr First-tier N° cases Aim of study
China 117 2012–2014 < 28 days 186 NB with hypotonia - Chromosome karyotyping 117 (22 peripheral hypotonia Evaluate speed and diagnostic
- LC-MS-MS disorders) specificity
- LS-PCR
- MLPA
- Sanger sequencing
- NGS

114
USA 2015–2017 <28 days 20 acutely ill infants Targeted gene panel sequencing 10 (1 congenital presynaptic Evaluate speed and
myasthenic syndrome 6) cost-effectiveness of the
diagnosis
India 116 N/A Fetus /NB 4 GSD IV (with severe WES 4 severe perinatally lethal Find novel pathogenic variants
neuromuscular form) neuromuscular forms of GSD
IV
120
Sweden N/A NB 1 hypotonic NB with WES Myopathy in Shwachman– Diagnosis
respiratory distress Diamond syndrome
113
USA 2021 –N/A <28 days 1 NB with hypotonia, WGS Neonatal-onset MYH2 hereditary Diagnosis
dysmorphic features, myosin myopathies
dysphagia
China 118 2019–2020 NB 93 NB small for gestational age - TMS 45 (4 Krabbe disease, 3 Examine the clinical application of
- Angel Care genomic screening cerebrotendinous genomic screening in
(panel of 150 diseases) xanthomatosis, 2 SMA, 1 newborns small for gestational
- tNGS primary carnitine deficiency) age

UEA 121 2021–2022 1–90 days 5 critically ill children Rapid WGS 3 (1 Diagnosis
dystrophy-dystroglycanopathy)
122
Belgium 2021–2022 2 days-18 21 critically ill children Rapid WGS 12 To establish the path for a
years nationwide semi-centered
rapid WGS network in
Belgium.
India 115 N/A NB 1 floppy NB Clinical exome sequencing Congenital hypomyelinating Diagnosis
neuropathy
South N/A NB 111 (55 NB tNGS 15 (1 congenital myasthenic Evaluate the possibility of rapid
Korea119 - hypotonia syndrome) and timely diagnosis of
- seizures treatable rare genetic diseases
- skeletal dysplasia)

List of abbreviations: LC-MS-MS: liquid chromatography coupled to tandem mass spectrometry; N° cases: Number of confirmed cases; N/A: not available; NB: newborn; NGS: next-generation sequencing; Pop:
description of the population screened; Ref: references used; Scr: Screening assay; tNGS: targeted next-generation sequencing; UEA: United Emirates Arab, WES: whole exome sequencing.
Journal of Neuromuscular Diseases 0(6)
Dangouloff et al. 11

symptoms affecting the brain, endocrine system, heart, units.114,117–119,122 In these studies, genetic testing used targeted
gastrointestinal tract, uterus, skin, and immune system panels or more extensive whole exome or whole genome
that differ based on age of onset. The disorder is caused sequencing with the goal of evaluating diagnostic speed and
by a pathological expansion of over 50 CTG repeats in accuracy. In the specific targeted populations, the diagnoses
the DMPK gene. MD1 onset occurs earlier with each suc- yield was between 27 and 63%, with peripheral hypotonia dis-
cessive generation, which typically happens with maternal orders, Krabbe disease, cerebrotendinous xanthomatosis,
transmission. The most severe form, congenital MD1, SMA, or congenital myasthenic syndrome identified Table 6.
occurs in 15% of cases. Patients present with severe gener-
alized weakness at birth, respiratory distress, hypotonia, and
feeding difficulties. Affected infants may experience Discussion
delayed cognitive and motor development, intellectual dis- Our literature search identified 87 articles describing 82 early
ability, and autism spectrum disorder, with the physical identification programs for neuromuscular diseases: 72
symptoms potentially leading to fatal outcomes. The inci- studies evaluated NBS programs for seven NMDs and ten
dence ranges from 0.5 to 1.8 per 100,000. studies focused on early genetic diagnosis for various diseases
Oligonucleotide-based agents and gene therapies have been including NMDs. The fast-growing number of peer-reviewed
tested in pre-clinical models and are currently in clinical devel- papers in this field illustrates the growing understanding of
opment. Nevertheless, no specific disease-modifying treat- the importance of early, and ideally pre-symptomatic, identifi-
ments are currently approved. Management primarily cation of patients with treatable conditions such as X-ALD,
involves monitoring for complications and providing assistive SMA, Pompe disease, and DMD. These diseases can be diag-
devices, hormone therapy, and pain management. nosed with a simple biochemical test or straightforward genetic
We identified an article describing a pilot project in the test. Congenital myasthenic syndrome is more difficult to diag-
USA that utilized deidentified dried blood spots to deter- nose, and we anticipate similar issues in limb-girdle muscular
mine the prevalence of MD1. The study found a prevalence dystrophies for which treatments are in development but no
rate of 1 in 21,100.112 Table 5 summarizes this article. simple and specific tests are available.
When there are no sensitive biochemical assays nor specific
hotspot mutations (such as the deletion of exon 7 of SMN1 in
Rapid genomic diagnosis patients with SMA), which is the case for the majority of
Exome and genome sequencing is increasingly used in neo- NMDs, screening for these disorders will not be compatible
natal intensive care units to diagnose and treat critically ill with the biochemical methods used in current NBS programs.
infants, but the gene panels used for these purposes are The fast-paced development of therapies also necessitates the
poorly reported and therefore little is known regarding the effi- ability to quickly add a gene to a genomic NBS program to
cacy of this type of testing. The conventional diagnostic ensure that patients are diagnosed as rapidly as possible.
approach based on hypotheses might not be effective in Some are hesitant about genomic NBS for several reasons.
these situations, as clinical manifestations during the neonatal One primary concern is the potential for false positives or uncer-
period are often nonspecific. Moreover, phenotypes linked to tain results, which can cause unnecessary stress and anxiety for
reduced survival in numerous genetic disorders may go unrec- families. Additionally, the interpretation of genetic data is
ognized due to early mortality. In subjects with NMDs, exome complex, and not all detected variants are fully understood,
and genome sequencing can enable diagnosis to be made. We leading to potential misdiagnoses or overdiagnoses. Enabling
identified articles reporting two pilot projects in the quicker diagnoses allows for more timely treatments, prevent-
USA,113,114 two in India,115,116 two in China,117,118 one in ing long-term complications and significantly improving the
South Korea,119 one in Sweden,120 one in the United effectiveness of interventions. For these reasons, several
Emirates Arab,121 and one in Belgium122 that evaluated use genomic NBS programs are currently underway that use
of genetic testing for diagnosis of NMDs. either using WGS or targeted next-generation sequencing.123
Five teams have reported unique cases diagnosed thanks Although the panels differ, all screen for mutations in genes
to genetic testing.113,115,116,120,121 These diagnoses enabled implicated in NMDs such as MLD. Parents appear positive
either early treatment, as was the case for patients with myop- about these first genomic NBS programs.124 As the cost of
athy in Shwachman-Diamond syndrome, a neonatal-onset adding a new disease is not a limitation when WGS is
MYH2 hereditary myosin myopathy, and congenital hypomye- employed, we expect that many NMDs will be added to the
linating neuropathy, or explained early death (as was the case list of diseases evaluated in NBS programs over the next few
for a patient with a severe perinatally lethal neuromuscular years. As both genomic NBS and early diagnostic testing will
form of glycogen storage disease IV).116 Five teams have tar- eventually utilize WGS, it is foreseeable that raw data of
geted specific populations: hypotonic children, children of WGS performed for NBS could be in the future re-analysed
low birth weight compared with gestational age, or children for diagnosis in a child presenting any serious condition.
with seizures, recruited from neonatal intensive care units, This scoping review has certain limitations. First, only
paediatric intensive care units and paediatric neurology one database (Medline) was consulted. We also restricted
12 Journal of Neuromuscular Diseases 0(6)

our search to original articles; conference presentations were

not considered. Data presented at neuromuscular disease Data availability
The data supporting the findings of this study are available within
conferences are likely to be published quickly, however, as
the article and/or its supplementary material.
the neuromuscular world is undergoing a period of transition
due to the recent approval of effective treatments.
References
Conclusion 1. Ciafaloni E, Fox DJ, Pandya S, et al. Delayed diagnosis in
duchenne muscular dystrophy: data from the Muscular
Since 2021, newborn screening for NMDs, notably in Dystrophy Surveillance, Tracking, and Research Network
X-ALD, SMA, and Pompe disease, have been incorporated (MD STARnet). J Pediatr 2009; 155: 380–385.
in routine NBS programs. Even for diseases where treatment 2. Lin C-W, Kalb SJ and Yeh W-S. Delay in diagnosis of
is currently not life-changing, such as Krabbe disease, new spinal muscular atrophy: a systematic literature review.
NBS programs continue to be implemented, especially in Pediatr Neurol 2015; 53: 293–300.
the USA. The use of genetic diagnostic tests does not yet 3. Kishnani PS, Amartino HM, Lindberg C, et al. Timing of diag-
appear to be widespread, or at least not widely reported. The nosis of patients with pompe disease: data from the pompe
genomic newborn screening programs currently underway registry. Am J Med Genet A 2013; 161A: 2431–2443.
will undoubtedly be a lever for rapid development. Rapid 4. van Geel BM, Assies J, Haverkort EB, et al. Delay in diag-
incorporation of tests into routine NBS will be critical as nosis of X-linked adrenoleukodystrophy. Clin Neurol
new treatments for NMDs become available. Neurosurg 1993; 95: 115–120.
5. Yang C-F, Yang CC, Liao H-C, et al. Very early treatment
Abbreviation for infantile-onset pompe disease contributes to better out-
DMD Duchenne muscular dystrophy comes. J Pediatr 2016; 169: 174–80.e1.
EMA European Medicines Agency 6. Dangouloff T and Servais L. Clinical evidence supporting early
FDA Food and Drug Administration treatment of patients with spinal muscular atrophy: current per-
MD1 Myotonic dystrophy 1 spectives. Ther Clin Risk Manag 2019; 15: 1153–1161.
MLD Metachromatic leukodystrophy
7. Mackels L and Servais L. The importance of early treatment
NBS Newborn screening
NMD neuromuscular disease of inherited neuromuscular conditions. J Neuromuscul Dis
RUSP Recommended Uniform Screening Panel 2024; 11: 253–274.
SMA spinal muscular atrophy 8. Delstanche S, Servais L and Gidaro T. Improved muscular
USA United States of America weakness during asthma exacerbation. JAMA Neurol 2017;
WGS whole genome sequencing
74: 353–354.
X-ALD X-linked adrenoleukodystrophy
9. Chae JH, Vasta V, Cho A, et al. Utility of next generation
sequencing in genetic diagnosis of early onset neuromuscu-
lar disorders. J Med Genet 2015; 52: 208–216.
Acknowledgments
10. Wilson JMG and Jungner G. Principles and practice of
The authors thank Jacky Wyatt for language editing. screening for disease. Public Health Paper, World Health
Organization 1968; 34: 11–13.
ORCID iD 11. Loeber JG, Platis D, Zetterström RH, et al. Neonatal screening
Tamara Dangouloff https://orcid.org/0000-0002-5435-4774 in Europe revisited: an ISNS perspective on the current state and
developments since 2010. Int J Neonatal Screen 2021; 7: 15.
Statements and declarations 12. Moher D, Liberati A, Tetzlaff J and Altman DG. Preferred
Funding reporting items for systematic reviews and meta-analyses:
The authors received no financial support for the research, author- the PRISMA statement. PLoS Med 2009; 6: e1000097.
ship, and/or publication of this article. 13. Crawford TO, Swoboda KJ, De Vivo DC, et al. Continued
benefit of nusinersen initiated in the presymptomatic stage
Declaration of conflicting interests of spinal muscular atrophy: 5-year update of the
The authors declared the following potential conflicts of interest NURTURE study. Muscle Nerve 2023; 68: 157–170.
with respect to the research, authorship, and/or publication of this 14. Strauss KA, Farrar MA, Muntoni F, et al. Onasemnogene
article: HL and NB have no conflicts of interest to report. TD has abeparvovec for presymptomatic infants with three copies
given lectures sponsored by Biogen and Roche. LS Servais has of SMN2 at risk for spinal muscular atrophy: the Phase III
given consultancy - attended the board of Zentech and Illumina. SPR1NT trial. Nat Med 2022; 28: 1390–1397.
Outside the scope of the paper, LS has given consultancy - attended 15. Strauss KA, Farrar MA, Muntoni F, et al. Onasemnogene
the board for Roche, Biogen, Novartis, Scholar Rock, BioHaven, abeparvovec for presymptomatic infants with two copies
Pfizer, PTC, Dyne, Wave, Biomarin, Myostana, MetrioPharma, of SMN2 at risk for spinal muscular atrophy type 1: the
Astellas, Sanofi, Sysnav, and RegenexBio. Phase III SPR1NT trial. Nat Med 2022; 28: 1381–1389.
Dangouloff et al. 13

16. Baranello G, Darras B, Day J, et al. Risdiplam in Type 1 Spinal results of a large-scale pilot project on 202,908 newborns.
Muscular Atrophy. N Engl J Med 2021; 384(10): 915–923. Pediatr Neurol 2024; 156: 147–154.
17. Aragon-Gawinska K, Moureaux C, Dangouloff T, et al. Spinal 33. Kiselev A, Maretina M, Shtykalova S, et al. Establishment
muscular atrophy treatment in patients identified by newborn of a pilot newborn screening program for spinal muscular
screening – a systematic review. Genes (Basel) 2023; 14: 1377. atrophy in Saint Petersburg. Int J Neonatal Screen 2024;
18. Weng W-C, Hsu Y-K, Chang F-M, et al. CMAP Changes 10: 9.
upon symptom onset and during treatment in spinal muscu- 34. Šimić D, Šarić A, Škarič ić A, et al. One-Year pilot study
lar atrophy patients: lessons learned from newborn screen- results of newborn screening for spinal muscular atrophy
ing. Genet Med 2021; 23: 415–420. in the Republic of Croatia. Int J Neonatal Screen 2024;
19. Kraszewski JN, Kay DM, Stevens CF, et al. Pilot study of 10: 50.
population-based newborn screening for spinal muscular 35. Brkušanin M, Garai N, Karanović J, et al. Our journey from
atrophy in New York state. Genet Med 2018; 20: 608–613. individual efforts to nationwide support: implementing
20. Vill K, Schwartz O, Blaschek A, et al. Newborn screening newborn screening for spinal muscular atrophy in Serbia.
for spinal muscular atrophy in Germany: clinical results Int J Neonatal Screen 2024; 10: 57.
after 2 years. Orphanet J Rare Dis 2021; 16: 53. 36. Boemer F, Caberg J-H, Beckers P, et al. Three years pilot of
21. D’Silva AM, Kariyawasam DST, Best S, et al. Integrating spinal muscular atrophy newborn screening turned into offi-
newborn screening for spinal muscular atrophy into health cial program in southern Belgium. Sci Rep 2021; 11: 19922.
care systems: an Australian pilot programme. Dev Med 37. Weng W-C, Hsu Y-K, Chang F-M, et al. CMAP Changes
Child Neurol 2021; 64: 625–632. upon symptom onset and during treatment in spinal muscu-
22. Kucera KS, Taylor JL, Robles VR, et al. A voluntary lar atrophy patients: lessons learned from newborn screen-
statewide newborn screening pilot for spinal muscular ing. Genet Med 2021; 23: 415–420.
atrophy: results from early check. Int J Neonatal Screen 38. Kraszewski JN, Kay DM, Stevens CF, et al. Pilot study of
2021; 7: 20. population-based newborn screening for spinal muscular
23. Abiusi E, Vaisfeld A, Fiori S, et al. Experience of a 2-year atrophy in New York state. Genet Med 2018; 20: 608–613.
spinal muscular atrophy NBS pilot study in Italy: towards 39. Vivo DD, Bertini E, Swoboda KJ, et al. Nusinersen initiated
specific guidelines and standard operating procedures for in infants during the presymptomatic stage of spinal muscu-
the molecular diagnosis. J Med Genet 2022; 60: 697–705. lar atrophy: interim efficacy and safety results from the
24. Kimizu T, Nozaki M, Okada Y, et al. Multiplex real-time phase 2 NURTURE study. Neuromuscular Disorders
PCR-based newborn screening for severe primary immuno- 2019; 29: 842–856.
deficiency and spinal muscular atrophy in Osaka, Japan: our 40. Abiusi E, Vaisfeld A, Fiori S, et al. Experience of a 2-year
results after 3 years. Genes (Basel) 2024; 15: 314. spinal muscular atrophy NBS pilot study in Italy: towards
25. Matteson J, Wu CH, Mathur D, et al. California’s experi- specific guidelines and standard operating procedures for
ence with SMA newborn screening: a successful path to the molecular diagnosis. J Med Genet 2023; 60: 697–705.
early intervention. J Neuromuscul Dis 2022; 9: 777–785. 41. Kernohan KD, McMillan HJ, Yeh E, et al. Ontario newborn
26. Kernohan KD, McMillan HJ, Yeh E, et al. Ontario newborn screening for spinal muscular atrophy: the first year. Can J
screening for spinal muscular atrophy: the first year. Can J Neurol Sci 2022; 49: 821–823.
Neurol Sci 2021; 49: 821–823. 42. Kay DM, Stevens CF, Parker A, et al. Implementation of
27. Olkhovych N, Gorovenko N and Servais L. Universal population-based newborn screening reveals low incidence
newborn screening for spinal muscular atrophy in of spinal muscular atrophy. Genetics in Medicine 2020; 8:
Ukraine. Lancet 2023; 402: 288–289. 1296–1302.
28. Sonehara S, Bo R, Nambu Y, et al. Newborn screening for 43. Dangouloff T, Boemer F, Caberg JH Correspondence on:
spinal muscular atrophy: a 2.5-year experience in Hyogo “discrepancy in spinal muscular atrophy incidence findings
Prefecture, Japan. Genes (Basel) 2023; 14: 2211. in newborn screening programs: the influence of carrier
29. Elkins K, Wittenauer A, Hagar AF, et al. Georgia State screening?” by Kay, et al. Genetics in Medicine 2020; 22:
spinal muscular atrophy newborn screening experience: 1913–1914.
screening assay performance and early clinical outcomes. 44. Dangouloff T, Vrscaj E, Servais L, et al. Newborn screening
Am J Med Genet C Semin Med Genet 2022; 190: 187–196. programs for spinal muscular atrophy worldwide: where we
30. Sawada T, Kido J, Sugawara K, et al. Newborn screening stand and where to go. Neuromuscul Disord 2021; 31: 574–582.
for spinal muscular atrophy in Japan: one year of experi- 45. Vršč aj E, Dangouloff T, Osredkar D, et al. Newborn screening
ence. Mol Genet Metab Rep 2022; 32: 100908. programs for spinal muscular atrophy worldwide in 2023.
31. Baker MW, Mochal ST, Dawe SJ, et al. Newborn screening J Neuromuscul Dis 2024. https://doi.org/10.1177/2214360
for spinal muscular atrophy: the Wisconsin first year experi- 2241288095.
ence. Neuromuscul Disord 2021; 32: 135–141. 46. Markati T, Oskoui M, Farrar MA, et al. Emerging therapies
32. Efimova IY, Zinchenko RA, Marakhonov A V, et al. for Duchenne muscular dystrophy. Lancet Neurol 2022; 21:
Epidemiology of spinal muscular atrophy based on the 814–829.
14 Journal of Neuromuscular Diseases 0(6)

47. Ellis JA, Vroom E and Muntoni F. 195th ENMC inter- 63. Chen H-A, Hsu R-H, Chen P-W, et al. High incidence of null
national workshop: newborn screening for duchenne mus- variants identified from newborn screening of X-linked adre-
cular dystrophy 14-16th December, 2012, naarden, The noleukodystrophy in Taiwan. Mol Genet Metab Rep 2022;
Netherlands. Neuromuscul Disord 2013; 23: 682–689. 32: 100902.
48. Scheuerbrandt G, Lundin A, Lövgren T, et al. Screening for 64. Tang C, Tang F, Cai Y, et al. A pilot study of newborn
duchenne muscular dystrophy: an improved screening test screening for X-linked adrenoleukodystrophy based on
for creatine kinase and its application in an infant screening liquid chromatography-tandem mass spectrometry method
program. Muscle Nerve 1986; 9: 11–23. for detection of C26:0-lysophosphatidylcholine in dried
49. Chung J, Smith AL, Hughes SC, et al. Twenty-year blood spots: results from 43,653 newborns in a southern
follow-up of newborn screening for patients with muscular Chinese pop. Clin Chim Acta 2024; 552: 117653.
dystrophy. Muscle Nerve 2016; 53: 570–578. 65. Bonaventura E, Alberti L, Lucchi S, et al. Newborn screen-
50. Moat SJ, Bradley DM, Salmon R, et al. Newborn bloodspot ing for X-linked adrenoleukodystrophy in Italy: diagnostic
screening for Duchenne muscular dystrophy: 21 years experi- algorithm and disease monitoring. Front Neurol 2023; 13:
ence in Wales (UK). Eur J Hum Genet 2013; 21: 1049–1053. 1072256.
51. Tavakoli NP, Gruber D, Armstrong N, et al. Newborn 66. Shimozawa N, Takashima S, Kawai H, et al. Advanced
screening for Duchenne muscular dystrophy: a two-year diagnostic system and Introduction of newborn screening
pilot study. Ann Clin Transl Neurol 2023; 10: 1383–1396. of adrenoleukodystrophy and peroxisomal disorders in
52. Drummond LM. Creatine phosphokinase levels in the Japan. Int J Neonatal Screen 2021; 7: 58.
newborn and their use in screening for Duchenne muscular 67. Albersen M, van der Beek SL, Dijkstra IME, et al.
dystrophy. Arch Dis Child 1979; 54: 362–366. Sex-specific newborn screening for X-linked adrenoleuko-
53. Plauchu H, Dorche C, Cordier MP, et al. Duchenne muscu- dystrophy. J Inherit Metab Dis 2023; 46: 116–128.
lar dystrophy: neonatal screening and prenatal diagnosis. 68. Lee S, Clinard K, Young SP, et al. Evaluation of X-linked
Lancet 1989; 1: 69. adrenoleukodystrophy newborn screening in North
54. Ke Q, Zhao Z-Y, Griggs R, et al. Newborn screening for Carolina. JAMA Netw Open 2020; 3: e1920356.
Duchenne muscular dystrophy in China: follow-up diagnosis 69. Matteson J, Sciortino S, Feuchtbaum L, et al.
and subsequent treatment. World J Pediatr 2017; 13: 197–201. Adrenoleukodystrophy newborn screening in California
55. Mendell JR, Shilling C, Leslie ND, et al. Evidence-based since 2016: programmatic outcomes and follow-up. Int J
path to newborn screening for Duchenne muscular dys- Neonatal Screen 2021; 7: 22.
trophy. Ann Neurol 2012; 71: 304–313. 70. Hall P, Li H, Hagar A, et al. Newborn screening for
56. Drousiotou A, Ioannou P, Georgiou T, et al. Neonatal screening X-linked adrenoleukodystrophy in Georgia: experiences
for Duchenne muscular dystrophy: a novel semiquantitative from a pilot study screening of 51,081 newborns. Int J
application of the bioluminescence test for creatine kinase in a Neonatal Screen 2020; 6: 81.
pilot national program in Cyprus. Genet Test 1998; 2: 55–60. 71. Priestley J, Adang L, Williams S, et al. Newborn screening
57. Greenberg CR, Rohringer M, Jacobs HK, et al. Gene for X-linked adrenoleukodystrophy: review of data and out-
studies in newborn males with Duchenne muscular comes in Pennsylvania. Int J Neonatal Screen 2022; 8: 24.
dystrophy detected by neonatal screening. Lancet 1988; 2: 72. Taylor JL and Lee S. Lessons learned from newborn screen-
425–427. ing in pilot studies. N C Med J 2019; 80: 54–58.
58. Kucera KS, Boyea BL, Migliore B, et al. Two years of 73. Baker C, Keller A, Lutz R, et al. Newborn screening for
newborn screening for duchenne muscular dystrophy as a X-linked adrenoleukodystrophy in Nebraska: initial experi-
part of the statewide early check research program in ences and challenges. Int J Neonatal Screen 2022; 8: 29.
North Carolina. Genet Med 2023; 26: 101009. 74. Burton B, Hickey R, Hitchins L, et al. Newborn screening
59. Jia C, Zhao D, Li Y, et al. Newborn screening and genomic for X-linked adrenoleukodystrophy: the initial Illinois
analysis of duchenne muscular dystrophy in henan, China. experience. Int J Neonatal Screen 2022; 8: 6.
Clin Chim Acta 2022; 539: 90–96. 75. Kemper AR, Brosco J, Comeau AM, et al. Newborn screen-
60. Chien Y-H, Lee N-C, Weng W-C, et al. Duchenne muscular ing for X-linked adrenoleukodystrophy: evidence summary
dystrophy newborn screening: the first 50,000 newborns and advisory committee recommendation. Genetics in
screened in Taiwan. Neurol Sci 2022; 43: 4563–4566. Medicine 2017; 19: 121–126.
61. Kucera KS, Boyea BL, Migliore B, et al. Two years of 76. Wiens K, Berry SA, Choi H, et al. A report on state-wide
newborn screening for Duchenne muscular dystrophy as a implementation of newborn screening for X-linked adreno-
part of the statewide early check research program in leukodystrophy. Am J Med Genet A 2019; 179: 1205–1213.
North Carolina. Genet Med 2024; 26: 101009. 77. Lee S, Clinard K, Young SP, et al. Evaluation of X-linked
62. Wiens K, Berry SA, Choi H, et al. A report on state-wide adrenoleukodystrophy newborn screening in North
implementation of newborn screening for X-linked Carolina. JAMA Netw Open 2020; 3: e1920356.
adrenoleukodystrophy. Am J Med Genet A 2019; 179: 78. Dhillon S. Avalglucosidase alfa: first approval. Drugs 2021;
1205–1213. 81: 1803–1809.
Dangouloff et al. 15

79. Luquetti DV, Jeng LJB, Donohue KM, et al. Regulatory mutations in the GAA gene in Japanese and Asian patients.
news: cipaglucosidase alfa-atga (Pombiliti) coadministered J Hum Genet 2019; 64: 741–755.
with Miglustat (Opfolda) for adults with late-onset pompe 96. Sawada T, Kido J, Sugawara K, et al. Current status of
disease. J Inherit Metab Dis 2024; 47: 578–581. newborn screening for Pompe disease in Japan. Orphanet
80. Yang C-F, Yang CC, Liao H-C, et al. Very early treatment J Rare Dis 2021; 16: 16.
for infantile-onset pompe disease contributes to better out- 97. Chang S, Zhan X, Liu Y, et al. Newborn screening for 6
comes. J Pediatr 2015; 169: 174–80.e1. lysosomal storage disorders in China. JAMA Netw Open
81. Lee N-C, Chang K-L, In ‘t Groen SLM, et al. Outcome of 2024; 7: e2410754.
later-onset pompe disease identified through newborn 98. Bravo H, Neto EC, Schulte J, et al. Investigation of new-
screening. J Pediatr 2022; 244: 139–147.e2. borns with abnormal results in a newborn screening
82. Yang C-F, Liao T-WE, Chu Y-L, et al. Long-term outcomes of program for four lysosomal storage diseases in Brazil.
very early treated infantile-onset pompe disease with short-term Mol Genet Metab Rep 2017; 12: 92–97.
steroid premedication: experiences from a nationwide newborn 99. Camargo Neto E, Schulte J, Pereira J, et al. Neonatal screen-
screening programme. J Med Genet 2023; 60: 430–439. ing for four lysosomal storage diseases with a digital micro-
83. Resources H, (HRSA) SA. Baby’s First Test 2020. fluidics platform: initial results in Brazil. Genet Mol Biol
84. Scott CR, Elliott S, Buroker N, et al. Identification of infants at 2018; 41: 414–416.
risk for developing Fabry, Pompe, or mucopolysaccharidosis-I 100. Kubaski F, Sousa I, Amorim T, et al. Pilot study of newborn
from newborn blood spots by tandem mass spectrometry. screening for six lysosomal diseases in Brazil. Mol Genet
J Pediatr 2013; 163: 498–503. Metab 2023; 140: 107654.
85. Wasserstein MP, Caggana M, Bailey SM, et al. The 101. Klug TL, Swartz LB, Washburn J, et al. Lessons learned
New York pilot newborn screening program for lysosomal from pompe disease newborn screening and follow-up. Int
storage diseases: report of the first 65,000 infants. Genet J Neonatal Screen 2020; 6: 11.
Med 2019; 21: 631–640. 102. Burton BK, Charrow J, Hoganson GE, et al. Newborn
86. Klug TL, Swartz LB, Washburn J, et al. Lessons learned screening for pompe disease in Illinois: experience with
from pompe disease newborn screening and follow-up. Int 684,290 infants. Int J Neonatal Screen 2020; 6: 4.
J Neonatal Screen 2020; 6: 11. 103. Orsini JJ, Escolar ML, Wasserstein MP, , et al. Krabbe
87. Ficicioglu C, Ahrens-Nicklas RC, Barch J, et al. Newborn Disease. In: MP Adam, HH Ardinger, RA Pagon,
Screening for Pompe Disease: Pennsylvania Experience. Int J SE Wallace, LJH Bean and K Stephens (eds)
Neonatal Screen 2020; 6: 89. GeneReviews® [Internet]. Seattle, WA: University of
88. Baerg MMM, Stoway SD, Hart J, et al. Precision newborn screen- Washington, Seattle, 1993–2020. 2000 Jun 19 [updated
ing for lysosomal disorders. Genet Med 2018; 20: 847–854. 2018 Oct 11], 1993.
89. Pillai NR, Fabie NAV, Kaye TV, et al. Disparities in late 104. Bradbury AM, Bongarzone ER and Sands MS. Krabbe
and lost: pediatricians’ role in following Pompe disease disease: new hope for an old disease. Neurosci Lett 2021;
identified by newborn screening. Mol Genet Metab 2023; 752: 135841.
140: 107633. 105. Wenger DA, Rafi MA, Luzi P, et al. Krabbe disease: genetic
90. Tang H, Feuchtbaum L, Sciortino S, et al. The first year aspects and progress toward therapy. Mol Genet Metab
experience of newborn screening for Pompe disease in 2000; 70: –9.
California. Int J Neonatal Screen 2020; 6: 9. 106. Orsini JJ, Kay DM, Saavedra-Matiz CA, et al. Newborn
91. Hall PL, Sanchez R, Hagar AF, et al. Two-tiered newborn screening for Krabbe disease in New York state: the first
screening with post-analytical tools for Pompe disease and eight years’ experience. Genet Med 2016; 18: 239–248.
mucopolysaccharidosis type I results in performance improve- 107. Basheeruddin K, Shao R, Balster F, et al. Newborn screen-
ment and future direction. Int J Neonatal Screen 2020; 6: 2. ing for Krabbe disease-Illinois experience: role of psycho-
92. WI State Laboratory of Hygiene. Pompe Pilot Conclusion. sine in diagnosis of the disease. Int J Neonatal Screen
Https://WwwWiaapOrg/Pompe-Pilot-Conclusion-Wi-State- 2021; 7: 24.
Laboratory-of-Hygiene/ 2019. 108. Ehmann P and Lantos JD. Ethical issues with testing and
93. Burlina AB, Polo G, Rubert L, et al. Implementation of second- treatment for Krabbe disease. Dev Med Child Neurol
tier tests in newborn screening for lysosomal disorders in North 2019; 61: 1358–1361.
Eastern Italy. Int J Neonatal Screen 2019; 5: 24. 109. Blackwell K, Gelb MH, Grantham A, et al. Family attitudes
94. Navarrete-Martínez JI, Limón-Rojas AE, de Jesús regarding newborn screening for Krabbe disease: results from
Gaytán-García M, et al. Newborn screening for six lysosomal a survey of leukodystrophy registries. Int J Neonatal Screen
storage disorders in a cohort of Mexican patients: three-year 2020; 6: 66.
findings from a screening program in a closed Mexican 110. Hong X, Daiker J, Sadilek M, et al. Toward newborn
health system. Mol Genet Metab 2017; 121: 16–21. screening of metachromatic leukodystrophy: results from
95. Momosaki K, Kido J, Yoshida S, et al. Newborn screening analysis of over 27,000 newborn dried blood spots. Genet
for Pompe disease in Japan: report and literature review of Med 2021; 23: 555–561.
16 Journal of Neuromuscular Diseases 0(6)

111. Wu THY, Brown HA, Church HJ, et al. Improving newborn 118. Zhang S, Zhou L, Zhang L, et al. Molecular genetic screen-
screening test performance for metachromatic leukodystro- ing of full-term small for gestational age. BMC Pediatr
phy: recommendation from a pre-pilot study that identified 2023; 23: 17.
a late-infantile case for treatment. Mol Genet Metab 2024; 119. Kim MJ, Kim SY, Lee JS, et al. Rapid targeted sequencing
142: 108349. using dried blood spot samples for patients with suspected
112. Johnson NE, Butterfield RJ, Mayne K, et al. actionable genetic diseases. Ann Lab Med 2023; 43: 280–289.
Population-Based prevalence of myotonic dystrophy type 120. Topa A, Tulinius M, Oldfors A, et al. Novel myopathy in a
1 using genetic analysis of statewide blood screening newborn with Shwachman-diamond syndrome and review
program. Neurology 2021; 96: e1045–e1053. of neonatal presentation. Am J Med Genet A 2016; 170A:
113. Oatmen K, Camelo-Piragua S and Zaghloul N. Novel muta- 1155–1164.
tion in the MYH2 gene in a symptomatic neonate with a her- 121. Halabi N, Ramaswamy S, El Naofal M, et al. Rapid whole
editary myosin myopathy. J Neonatal Perinatal Med 2022; genome sequencing of critically ill pediatric patients from
15: 63–68. genetically underrepresented populations. Genome Med
114. Brunelli L, Jenkins SM, Gudgeon JM, et al. Targeted gene 2022; 14: 56.
panel sequencing for the rapid diagnosis of acutely ill 122. Lumaka A, Fasquelle C, Debray FG, et al. Rapid whole
infants. Mol Genet Genomic Med 2019; 7: e00796. genome sequencing diagnoses and guides treatment in crit-
115. Ravikumar S, Devi U, Balakrishnan U, et al. Diagnosis of a ically Ill children in Belgium in less than 40 hours. Int J Mol
floppy neonate with misleading clues: unraveled as congeni- Sci 2023; 24: 400.
tal hypomyelinating neuropathy. BMJ Case Rep 2022; 15: 123. Minten T, Gold NB, Bick S, et al. Determining the charac-
e247555. teristics of genetic disorders that predict inclusion in
116. Radhakrishnan P, Moirangthem A, Nayak SS, et al. Novel newborn genomic sequencing programs. MedRxiv 2024:
pathogenic variants in GBE1 causing fetal akinesia deform- 2024.03.24.24304797. https://doi.org/10.1101/2024.03.24.
ation sequence and severe neuromuscular form of glycogen 24304797.
storage disease type IV. Clin Dysmorphol 2019; 28: 17–21. 124. Dangouloff T, Hovhannesyan K, Mashhadizadeh D, et al.
117. Wang Y, Peng W, Guo H-Y, et al. Next-generation Feasibility and acceptability of a newborn screening program
sequencing-based molecular diagnosis of neonatal hypo- using targeted next-generation sequencing in one maternity
tonia in Chinese population. Sci Rep 2016; 6: 29088. hospital in southern Belgium. Children 2024; 11: 26.