Instruction for Use

gastroplexVirus
real time RT-PCR Kit
For the qualitative in-vitro detection of RNA from Rotavirus, Norovirus (GI and
GII), and DNA from Adenovirus in clinical specimens, environmental and food
samples.

G01086-32 G01086-96

32 96

gerbion gmbH & Co. KG

Remsstr. 1
70806 Kornwestheim
Germany
phone: +49 7154 806 20 0
fax: + 49 7154 806 20 29
e-mail: [email protected]
www.gerbion.com

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Index
1 Intended Use ............................................................................................................................................. 3
2 Pathogen Information ........................................................................................................................... 3
3 Principle of the Test ............................................................................................................................... 4
4 Package Contents ................................................................................................................................... 5
5 Equipment and Reagents to be Supplied by User ................................................................... 5
6 Transport, Storage and Stability....................................................................................................... 5
7 Important Notes ...................................................................................................................................... 6
8 General Precautions ............................................................................................................................... 6
9 Sample Material ...................................................................................................................................... 6
10 Sample Preparation ............................................................................................................................... 6
11 Control RNA ................................................................................................................................................ 7
12 Real time RT-PCR .................................................................................................................................... 8
12.1 Important Points Before Starting:....................................................................................... 8
12.2 Procedure ....................................................................................................................................... 8
13 Instrument Settings............................................................................................................................. 10
14 Data Analysis ......................................................................................................................................... 12
15 Assay Validation ................................................................................................................................... 14
16 Limitations of the Method ............................................................................................................... 14
17 Troubleshooting .................................................................................................................................... 14
18 Kit Performance .................................................................................................................................... 16
18.1 Diagnostic Sensitivity and Specificity ............................................................................ 16
18.2 Analytical Sensitivity ............................................................................................................. 16
18.3 Analytical Specificity ............................................................................................................. 18
19 Abbreviations and Symbols ............................................................................................................. 19
20 Literature ................................................................................................................................................. 19

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1 Intended Use
The gastroplexVirus real time RT-PCR is an assay for the detection of the RNA
of Rotavirus, Norovirus, and the DNA of Adenovirus in clinical specimens (e.g.
stool samples, vomit), environmental and food samples using real time PCR
microplate systems.
2 Pathogen Information
Acute gastroenteritis is a worldwide major cause of morbidity and mortality.
Gastroenteritis or infectious diarrhea is an inflammation of the gastrointestinal
tract. Both the stomach and the small intestine are involved. Typical symptoms
are diarrhoea, vomiting, abdominal pain, and cramps, often followed by
dehydration.
The causative agent can be viral or bacterial. Enteric viruses are the major
pathogens for gastroenteritis especially in children. Noro-, Rota-, Adeno-, Sapo-
and Astroviruses are the most important viral pathogens.

Noroviruses are small non-enveloped RNA viruses belonging to the family of

Caliciviridae. They cause approximately 90 % of epidemic non-bacterial
outbreaks of gastroenteritis around the world.
The viruses are transmitted by fecally contaminated food or water and by
person-to-person contact. For this reason, outbreaks of norovirus infection
often occur in closed or semi-closed communities, such as long-term care
facilities, hospitals, prisons, dormitories, and cruise ships. Noroviruses are highly
contagious and are stable at temperatures between -20°C to +60°C and in
acidic environments up to pH 3. Norovirus infections occur throughout the year,
however, in Europe, seasonal increases are observed between October and
March.
The gastroplexVirus real time RT-PCR detects Norovirus strains of high genetic
diversity, such as the following:
GI: Norwalk, Desert Shield, Winchester, Queensarms, Southhampton, Chiba
GII: Lordsdale, Bristol, Melksham, Toronto, Hawaii
Infections with Rotavirus are the most common cause of severe diarrhoea
among children. Worldwide more than 450,000 children under 5 years of age
die from rotavirus infections each year. Most of them in developing countries.
The double-stranded RNA virus of the family Reoviridae is transmitted faecal-
orally and infects the enterocytes. It causes diarrhoea, vomiting, fever, and
dehydration, seldomly abdominal pain. Sometimes infections of the upper

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respiratory tract occur in correlation with gastroenteritis. With each infection

immunity develops, so subsequent infections are less severe. By the age of 5,
nearly every child in the world has at least once gone through a rotavirus
infection.
Rotaviruses are classified into the groups A-G, among which A-C are human
pathogenic. More than 90 % of rotavirus infections are caused by group A
viruses.

Adenoviruses mainly cause infections of the respiratory system. However,

dependent on the serotype, numerous other diseases can be caused, such as
gastroenteritis, keratoconjunctivitis epidemica, cystitis, rhinitis, pharyngitis, and
diarrhoea. Respiratory symptoms range from mild flu to acute bronchitis and
pneumonia. Immuno-suppressed patients are prone to severe complications,
such as acute respiratory distress syndrome. Although the epidemiological
characteristics of Adenoviruses vary from type to type, all types are
transmitted by direct contact, feacal-orally, and rarely by water. Some types
cause persistent, asymptomatic infections of the palatine and pharyngeal
tonsils, and the gastrointestinal tract. Spreading of the virus can occur over
months or years.
3 Principle of the Test
The gastroplexVirus real time RT-PCR contains specific primers and hydrolysis
probes for the detection of the nucleic acids of Rotavirus, Adenovirus, and
Norovirus in clinical specimens (e.g. stool samples, vomit), environmental and
food samples. The reverse transcription (RT) of viral RNA to cDNA and the
subsequent amplification of virus specific fragments are performed in a one-
step RT-PCR. The amplification can be detected when specific probes are
hydrolysed by the Polymerase. The emitted fluorescence is measured in the
FAM (Norovirus), ROX (Rotavirus) and Cy5 (Adenovirus) channel.
Furthermore, the gastroplexVirus real time RT-PCR contains a Control RNA,
which is detected in a second amplification system. Added during RNA
extraction, the Control RNA allows not only for the detection of RT-PCR
inhibition but also detects possible mistakes during RNA extraction. This greatly
reduces the risk of false-negative results. The Control RNA can also be used
solely as Internal Control by adding it directly to the Mastermix. The
fluorescence of the Control RNA is measured in the VIC®/HEX/JOETM/TET
channel.

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4 Package Contents
The reagents supplied are sufficient for 32 or 96 reactions respectively.
Table 1: Components of gastroplexVirus real time RT-PCR kit.
Content
Label Lid Colour
32 96
Reaction Mix yellow 1 x 506 µl 2 x 759 µl
Enzyme blue 1 x 6.4 µl 1 x 19.2 µl
Positive Control red 1 x 50 µl 1 x 100 µl
Negative Control green 1 x 50 µl 1 x 100 µl
Control RNA colourless 1 x 160 µl 2 x 240 µl

5 Equipment and Reagents to be Supplied by User

• RNA isolation kit (e.g. NukEx Pure RNA/DNA or NukEx Mag RNA/DNA)
• PCR grade Water
• Sterile microtubes
• Pipets (adjustable volume)
• Sterile pipet tips with filter
• Table centrifuge
• Vortexer
• Real time PCR instrument
• Optical PCR reaction tubes with lid
• Optional: Liquid handling system for automation
• Optional: VLP-RNA (please look at page 7 for details)
6 Transport, Storage and Stability
gastroplexVirus real time RT-PCR is shipped on dry ice or cool packs. All
components must be stored at maximum -18°C in the dark immediately after
receipt. Do not use reagents after the date of expiry printed on the package.
Up to 20 freeze and thaw cycles are possible. For convenience, opened
reagents can be stored at +2-8°C for up to 6 months.
Protect kit components from direct sunlight during the complete test run.

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7 Important Notes
• gastroplexVirus real time RT-PCR must be performed by qualified personnel
only.
• Good Laboratory Practice (GLP) has to be applied.
• Clinical samples must always be regarded as potentially infectious material
and all equipment used has to be treated as potentially contaminated.
8 General Precautions
• Stick to the protocol described in the Instruction for use.
• Set up different laboratory areas for the preparation of samples and for the
set up of the RT-PCR in order to avoid contaminations.
• Pipettes, tubes and other materials must not circulate between those
different laboratory areas.
• Always use filter tips.
• Regulary decontaminate equipment and benches with ethanol-free
decontaminant.
• Do not combine gastroplexVirus real time RT-PCR components of different
lot numbers.
9 Sample Material
Starting material for the gastroplexVirus real time RT-PCR is the nucleic acid
isolated from clinical specimens (e.g. stool samples, vomit), environmental or
food samples.
10 Sample Preparation
The gastroplexVirus real time RT-PCR is suitable for the detection of Rotavirus,
Adenovirus, and Norovirus in clinical specimens (e.g. stool samples, vomit),
environmental and food samples isolated with suitable isolation methods.
Commercial kits for RNA isolation such as the following are recommended:
• NukEx Pure RNA/DNA
• NukEx Mag RNA/DNA

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Important:
In addition to the samples always run a ‚water control‘ in your extraction. Treat
this water control analogous to a sample.
Comparing the amplification of the Control RNA in the samples to the
amplification of the internal control in the water control will give insights on
possible inhibitions of the real time RT-PCR. Furthermore, possible
contaminations during RNA extraction will be detectable.

Please note the chapter 11 ‚Control RNA‘.

If the real time RT-PCR is not performed immediately, store extracted RNA and
DNA according to the instructions given by the manufacturer.
11 Control RNA
A Control RNA is supplied and can be used as extraction control or only as
inhibition control. This allows the user to control the RNA isolation procedure
and to check for possible real time RT-PCR inhibition.
The Virus-Like Particles (VLP-RNA) are not supplied, but must be added to the
clinical or environmental samples directly. VLP-RNA can be used as patient-
side extraction control and in automated extraction systems, when pipetting of
the Control RNA to the first buffer of (e.g. binding buffer) of the respective
extraction kit is not possible due to extraction instrument specifications.

RNA isolation from clinical specimens (e.g. stool samples, vomit),

environmental and food samples

a) Control RNA or VLP-RNA used as Extraction Control:

gastroplexVirus real time RT-PCR Control RNA or VLP-RNA is added to the RNA
extraction.
Add 5 µl Control RNA or VLP-RNA per extraction (5 µl x (N+1)). Mix well.
Perform the RNA isolation according to the manufacturer‘s instructions. Please
follow protocol A.

The Control RNA must be added to the Lysis Buffer of the extraction kit.
b) Control RNA used as Internal Control of the real time PCR:
If only inhibition will be checked please follow protocol B.

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12 Real time RT-PCR

12.1 Important Points Before Starting:
• Please pay attention to the chapter 7 ‚Important Notes‘.
• Before setting up the real time RT-PCR familiarise yourself with the real
time PCR instrument and read the user manual supplied with the
instrument.
• The programming of the thermal profile should take place before the RT-
PCR set up.
• In every RT-PCR run one Positive Control and one Negative Control should
be included.
• Before each use, all reagents - except the Enzyme - should be thawed
completely at room temperature, thoroughly mixed (do NOT vortex the
Reaction Mix but mix by pipetting up and down repeatedly), and centrifuged
very briefly.

12.2 Procedure
If the Control RNA or VLP-RNA is used to control both, the real time RT-PCR and
the RNA isolation procedure, please follow protocol A. If the Control RNA is
solely used to detect possible inhibition of the real time RT-PCR, please follow
protocol B.
Protocol A
The Control RNA or VLP-RNA was added during RNA extraction (see chapter 11
‚Control RNA‘). In this case, prepare the Master Mix according to Table 2.
The Master Mix contains all of the components needed for RT-PCR except the
sample. Prepare a volume of Master Mix for at least one sample more than
required, in order to compensate for pipetting inaccuracy.
Table 2: Preparation of the Master Mix (Control RNA or VLP-RNA was added
during RNA extraction)

Volume per Reaction Volume Master Mix

15.8 µl Reaction Mix 15.8 µl x (N+1)

0.2 µl Enzyme 0.2 µl x (N+1)

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Protocol B
The Control RNA is used for the control of the real time RT-PCR only (see
chapter 11 ‚Control RNA‘). In this case, prepare the Master Mix according to
Table 3.
The Master Mix contains all of the components needed for real RT-PCR except
the sample. Prepare a volume of Master Mix for at least one sample more than
required, in order to compensate for pipetting inaccuracy.

Important: Dilute the Control RNA 1:10 in PCR grade Water (e.g. 1 µl Control
RNA + 9 µl PCR grade Water) before adding it to the Master Mix.
Table 3: Preparation of the Master Mix (Control RNA is added directly to the
Master Mix)

Volume per Reaction Volume Master Mix

15.8 µl Reaction Mix 15.8 µl x (N+1)

0.2 µl Enzyme 0.2 µl x (N+1)

0.2 µl Control RNA* (diluted 1:10) 0.2 µl x (N+1)*

*The increase in volume caused by adding the Control RNA is not taken into account
when preparing the PCR assay.

Protocol A and B: real time RT-PCR set up

• Place the number of optical PCR reaction tubes needed into the respective
tray of the real time PCR instrument.
• Pipet 16 µl of the Master Mix into each optical PCR reaction tube.
• Add 4 µl of the eluates from the RNA isolation (including the eluate of the
water control), the Positive Control and the Negative Control to the
corresponding optical PCR reaction tube (Table 4).
• Close the optical PCR reaction tubes immediately after filling in order to
reduce the risk of contamination.

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Table 4: Preparation of the real time PCR

Component Volume

Master Mix 16.0 µl

Sample 4.0 µl

Total Volume 20.0 µl

12.3 Instrument Settings

For the real time RT-PCR use the thermal profile shown in table 5.
Table 5: real time RT-PCR thermal profile
Description Time Temperature Number of
Cycles

Reverse Transcription 10 min 45°C 1

Initial Denaturation 5 min 95°C 1
Amplification of cDNA
Denaturation 10 sec 95°C
45
Annealing and Extension 40 sec 60°C
Aquisition at the end of this step

Dependent on the real time instrument used, further instrument settings have
to be adjusted according to table 6.

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Table 6: Overview of the instrument settings required for gastroplexVirus real

time RT-PCR.

Real time PCR Detection

Instrument Parameter Channel Notes
Norovirus 483-533 gerbion LC480 Colour
Rotavirus 558-610 Compensation required
LightCycler 480I
Control RNA 523-568
Adenovirus 615-670
gerbion LC480 Colour
Compensation required
Max
Melt Quant
Integration
Factor Factor
Time (sec)
LightCycler 480II
Norovirus 465-510 1 10 1
Rotavirus 533-610 1 10 2
Control RNA 533-580 1 10 2
Adenovirus 618-660 1 10 3
Norovirus FAM Gain 8
Stratagene
Rotavirus ROX Gain 1 Reference
Mx3000P /
Control RNA HEX Gain 1 Dye: None
Mx3005P
Adenovirus Cy5 Gain 4
Norovirus FAM
Rotavirus ROX
ABI 7500 Option Reference Dye ROX: NO
Control RNA JOE
Adenovirus Cy5
Norovirus Green Gain 5
Rotor-Gene Q,
Rotavirus Orange Gain 5
Rotor-Gene 3000
Rotor-Gene 6000 Control RNA Yellow Gain 5
Adenovirus Red Gain 5

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13 Data Analysis
The virus specific amplifications are measured in the FAM, ROX, Cy5 channels.
The amplification of the Control RNA is measured in the VIC®/HEX/JOE/TET
channel. The Positive Control contains in vitro transcripts of the respective
nucleic acid sequences of Rotavirus, Norovirus and Adenovirus. For the Positive
Control signals in the FAM, ROX, Cy5 channels must be detected.
Following results can occur:
• A signal in the FAM, ROX, Cy5 channels is detected:
The result is positive, the sample contains viral RNA.
In this case, detection of a signal of the Control RNA in the VIC®/HEX/JOE/TET
channel is inessential, as high concentrations of cDNA may reduce or
completely inhibit amplification of the Control RNA.
• No signal in the FAM, ROX, Cy5 channels, but a signal in the
VIC®/HEX/JOE/TET channel is detected:
The result is negative, the sample does not contain viral RNA.
The signal of the Control RNA excludes the possibilities of RNA isolation
failure (in case the Control RNA is being used as an Extraction Control)
and/or real time RT-PCR inhibition. If the CT value of a sample differs
significantly from the CT value of the water control, a partial inhibition
occured, which can lead to negative results in weak positive samples (see
‘Troubleshooting’).
• Neither in the FAM, ROX, Cy5 channels nor in the VIC®/HEX/JOE/TET channel
a signal is detected:
A diagnostic statement cannot be made.
The RNA isolation was not successful or an inhibition of the RT-PCR has
occurred. In case the Control RNA was added during RNA isolation and not
directly to the PCR Master Mix, the Negative Control is negative in both
channels.

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Figure 1 and Figure 2 show examples for positive and negative real time RT-
PCR results.

Figure 1: The positive sample shows virus-specific amplification in the FAM

channel, whereas no fluorescence signal is detected in the negative sample.

Figure 2: The positive sample as well as the negative sample show a signal in
the Control RNA specific VIC®/HEX/JOE/TET channel. The amplification signal of
the Control RNA in the negative sample shows, that the missing signal in the
virus specific FAM channel is not due to RT-PCR inhibition or failure of RNA
isolation, but that the sample is a true negative.

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14 Assay Validation
Set a threshold as follows:

Negative Controls
All negative controls should be below the threshold. If there is a potential
contamination (appearance of a curve in the negative control or a cluster of
curves in specimens at high CT – for example above 36), results obtained are
not interpretable and the whole run (including extraction) has to be repeated.

Positive Controls
All the positive controls must show a positive (i.e. exponential) amplification
curve. The positive controls must fall below a CT of 30.

Internal Controls
All internal controls must show a positive (i.e. exponential) amplification curve.
The internal control must fall below a CT of 33. If the internal control is above
CT 34, this points to a purification problem or a strong positive sample that can
inhibit the IC. In the latter case, the assay is valid. If a water control run is
performed, the IC must fall below a CT of 33.
15 Limitations of the Method
The results must always be considered in relation to the clinical symptoms.
Therapeutical consequences should be made in consideration of clinical data.
A negative test result does not exclude a gastroenteric virus infection.
16 Troubleshooting
The following troubleshooting guide is included to help you with possible
problems that may arise when performing a real time RT-PCR.

No fluorescence signal in the FAM, ROX, Cy 5 channel of the Positive Controls

The selected channel for Select the FAM channel for analysis of the Norovirus
analysis does not comply specific amplification, the ROX channel for analysis of
with the protocol the Rotavirus specific amplification and the Cy5
channel for analysis of the Adenovirus specific
amplification (Reaction Mix). Select the
VIC®/HEX/JOETM/TET cannel for the amplification of the
Control RNA. Due to amplification in the specific
channels, amplification of the Internal Control can be
inhibited in the Positive Control.
Incorrect configuration of Check your work steps and compare with ‚Procedure‘
the real time RT-PCR on page 8.

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The programming of the Compare the thermal profile with the protocol (Table
thermal profile is incorrect 5, page 10).
Incorrect storage conditions Check the storage conditions and the date of expiry
for one or more kit printed on the kit label. If necessary, use a new kit and
components or kit expired make sure kit components are stored as described in
‚Transport, Storage and Stability‘, page 5.
Weak or no signal of the Control RNA and simultaneous absence of a signal in the
virus specific FAM channel, ROX channel or Cy5 channel.
real time RT-PCR conditions Check the real time RT-PCR conditions (page 8).
do not comply with the
protocol
real time RT-PCR inhibited Make sure that you use an appropriate isolation
method (see ‚Sample Preparation‘, page 6) and follow
the manufacturer‘s instructions. Make sure that the
ethanol-containing washing buffer of the isolation kit
has been completely removed. An additional
centrifugation step at high speed is recommended
before elution of the RNA.
RNA loss during isolation In case the Control RNA was added before extraction,
process the lack of an amplification signal can indicate that
the RNA isolation was not successful. Make sure that
you use an appropriate isolation method (commercial
kits are recommended) and stick to the
manufacturer‘s protocol.
Incorrect storage conditions Check the storage conditions and the date of expiry
for one or more printed on the kit label. If necessary, use a new kit and
components or kit expired make sure kit components are stored as described in
‚Transport, Storage and Stability‘, page 5.
Detection of a fluorescence signal in the FAM channel, ROX channel or Cy5 channel
of the Negative Control.
Contamination during Repeat the real time RT-PCR in replicates. If the result
preparation of the RT-PCR is negative in the repetition, the contamination
occured when the samples were pipetted into the
optical PCR reaction tubes. Make sure to pipet the
Positive Control last and close the optical PCR reaction
tube immediately after adding the sample. If the
same result occurs, one or more of the kit
components might be contaminated. Make sure that
work space and instruments are decontaminated
regularly. Use a new kit and repeat the real time RT-
PCR.
Detection of a weak fluorescence signal in the FAM channel of a sample with a
strong fluorescence signal in the Cy5 channel.
Cross-talk Depending on the real time PCR instrument used, a
strong fluorescence signal in one detection channel
can lead to a weak signal (around CT 40) in another
channel due to so-called cross-talk between channels.

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17 Kit Performance
17.1 Diagnostic Sensitivity and Specificity
During the validation study of the gastroplexVirus real time RT-PCR 163
positive and 89 samples, negative for enteric pathogens were tested. The
positive samples were also tested for the other pathogens to exclude
unspecific reactions. The diagnostic sensitivity was found to be 100 % and the
diagnostic specificity 100 %.
The positive predictive value was found to be 100 %, the negative predictive
value showed to be 100 %.

Table 7: Overview of the amount of samples tested and the resulting

sensitivity and specificity.
gastroplexVirus
gastroplexVirus positive
negative
Norovirus positive 45 0
Norovirus negative 0 207
Rotavirus positive 39 0
Rotavirus negative 0 213
Adenovirus positive 35 0
Adenovirus negative 0 217
Sensitivity 100 %
Specificity 100 %

17.2 Analytical Sensitivity

The limit of detection (LoD) of gastroplexVirus real time RT-PCR was
determined using serial dilutions of in vitro transcripts (RNA Viruses) and cloned
DNA (Adenovirus) in nucleic acid stabilization buffer in a Stratagene Mx3000
real time PCR instrument. Total nucleic acids were extracted using NukEx Pure
RNA/DNA according to the manufacturer´s instructions. Each sample (200 µl of
diluted nucleic acid) was supplemented with 5 µl Control-RNA prior to
extraction. Total nucleic acids were eluted with 50 µl and 4 µl of the eluates
were applied to the subsequent real time RT-PCR.
The LoD of gastroplexVirus real time RT-PCR for Norovirus, Rotavirus, and
Adenovirus is >10 copies per reaction each.

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Table 8: Samples tested for the validation of the sensitivity of gastroplexVirus.

Copies per Expected CT-value
Norovirus Mean CT
Reaction Result gastroplexVirus
Noro 10-2 1.000.000 positive 20.20/ 20.52 20.36
Noro 10-3 100.000 positive 23.65/ 23.12 23.39
Noro 10-4 10.000 positive 26.44/ 26.69 26.57
Noro 10-5 1.000 positive 30.16/ 29.81 29.98
Noro 10-6 100 positive 32.4/ 33.36 32.88
Noro 10-7 10 positive 36.27/ 35.82 36.05
Noro 10-8 1 positive 39.73/ no Ct 39.73

Copies per Expected CT-value

Rotavirus Mean CT
Reaction Result gastroplexVirus
Rota 10-2 1.000.000 positive 20.43/ 20.74 20.59
Rota 10-3 100.000 positive 23.27/ 23.19 23.23
Rota 10-4 10.000 positive 26.89/ 27.22 27.10
Rota 10-5 1.000 positive 31.56/ 31.71 31.64
Rota 10-6 100 positive 34.84/ 35.18 35.01
Rota 10-7 10 positive 38.41/ 38.62 38.52
Rota 10-8 1 positive no Ct/ no Ct -

Copies per Expected CT-value

Adenovirus Mean CT
Reaction Result gastroplexVirus
Adeno 10-2 1.000.000 positive 20.49/ 20.05 20.27
Adeno 10-3 100.000 positive 23.58/ 23.29 23.44
Adeno 10-4 10.000 positive 26.95/ 27.42 27.19
Adeno 10-5 1.000 positive 31.12/ 31.5 31.31
Adeno 10-6 100 positive 34.34/ 34.59 34.47
Adeno 10-7 10 positive 37.79/ 36.88 37.34
Adeno 10-8 1 positive 39.86/ 40.42 40.14

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17.3 Analytical Specificity

The specificity of gastroplexVirus real time RT-PCR was evaluated by in silico
analysis and additionally by amplification of RNA and DNA of other relevant
viruses and bacteria found in clinical samples.
Results:
The gastroplexVirus real time RT-PCR showed a positive result for the samples
containing Noroviruses, Rotavirus, and Adenovirus, whereas samples
containing other pathogens were reliably tested negative. The results are
shown in Table 9.
Table 9: Bacterial and viral pathogens tested for the determination of the
analytical specificity of gastroplexVirus real time RT-PCR.
Strain Expected Result Result
Enterovirus 68 negative negative
Coxsackievirus B3 negative negative
Coxsackievirus A16 negative negative
Coxsackievirus B5 negative negative
Salmonella negative negative
Listeria monocytogenes negative negative
Escherichia coli negative negative
Campylobacter negative negative
Shigella negative negative
Yersinia negative negative
Sapovirus negative negative
Astrovirus negative negative
Norovirus GI positive positive
Norovirus GII positive positive
Rotavirus positive positive
Adenovirus positive positive

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18 Abbreviations and Symbols

cDNA complementary
Catalog number
Deoxyribonucleid Acid
RNA Contains sufficient for
Ribonucleid Acid
<n> test
PCR Polymerase Chain Upper limit of
Reaction temperature
RT Reverse Transcription Manufacturer
REACTION MIX Reaction Mix Use by YYYY-MM
ENZYME Enzyme Batch code
CONTROL Positive Control Content
Consult instructions for
CONTROL Negative Control
use
CONTROL RNA IC
In vitro diagnostic
Control RNA
medical device

19 Literature
[1] Lothar Thomas, Labor und Diagnose: Indikation und Bewertung von
Laborbefunden für die medizinische Diagnostik, 8. Auflage, 2012, TH-Books,
ISBN-10: 3980521583

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