53

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112:15701576.
 62

Lampiran 1. Ethical Clearance

 63

Lampiran 2. Prosedur Analisis Sitogenetika

1. Alat

- Spuit 3cc untuk mengambil sampel darah pasien

- Gelas ukur

- Tabung steril 15 ml

- Tabung vacuum 3cc

- Tube eppendrofe 1,5 ml

- Blue tip, yellow tip

- Pipet pastur steril

- Sentrifuse

- Inkubator

- Waterbath

- Almari es

- Almari asam

- Object glass

- Rak tabung reaksi

- Botol semprot untuk KCl

- Tissue

- Laminar flow

- Mikroskop cahaya, mikroskop fotografi

2. Bahan

Bahan yang diperiksa adalah darah vena dengan antikoagulan heparin.

 64

3. Reagen yang dibutuhkan

- Media kultur kromosom

Media yang digunakan adalah MEM dan RPMI 1640

- Bahan pemicu mitosis yaitu Phytohaemaglutinin-mixture (PHA-M)

- Fetal bovine Serum (FBS) 10% dari kadar medium sebagai nutrisi

- Colcemid untuk menghentikan mitosis (spindle inhibitor)

- KCl 0,075 M sebagai larutan hipotonik untuk memecah sel

- Larutan Carnoys (3 metanol : 1 asam asetat glacial) sebagai larutan fiksasi

sel

- Giemsa

- Buffer fosfat

- Phosphat Buffer Saline (PBS) pH 6,8

- Larutan Tripsin 1% stok dalam PBS

- Aquabides

- Immersion Oil

1. Prosedur Pengambilan Bahan

Sebelum mengambil darah pasien, dicatat identitas pasien, diberi informed

consent, dan pemeriksaan fisik terlebih dahulu. Sampel darah diambil dari

pembuluh darah vena cubiti dengan menggunakan antikoagulan heparin.

 65

2. Prosedur Pemeriksaan Bahan

- Penanaman

1. Mengeluarkan medium, PHA-M, dan FBS dari almari es dan

dihangatkan dalam waterbath 37C selama 15 menit.

2. Meneteskan FBS sebanyak 10% dari jumlah medium kedalam masing-

masing tabung berisi media MEM dan RPMI 1640, lalu dicampur

secara perlahan.

3. Menambahkan 100l PHA-M kedalam tabung dan dicampur kembali

secara perlahan.

4. Jika sampel darah baru diambil, proses penanaman dilanjutkan dengan

meneteskan 4 tetes buffy coat dan 6 tetes whole blood kedalam tabung.

Namun, jika sampel darah sudah lama (sampel darah sudah didiamkan

pada suhu 4C) penanaman dilanjutkan dengan meneteskan 10 tetes

whole blood kedalam tabung. Selanjutnya, tabung dikocok lagi secara

perlahan-lahan.

5. Melakukan inkubasi sampel pada suhu 37 C selama 72-96 jam dengan

sudut kemiringan tabung 45 agar memberi peluang tumbuhnya sel di

permukaan, dalam inkubator.

6. Setiap 1x24 jam, masing-masing tabung dikocok secara perlahan-

lahan.

*Langkah 2, 3, dan 4 dilakukan dalam laminar flow.

 66

- Pemanenan

1. Meneteskan 100 l Colcemid pada setiap tabung, dan diinkubasi dalam

inkubator pada suhu 37 selama 30 menit.

2. Setelah dilakukan inkubasi, lalu dipusingkan selama 10 menit dengan

kecepatan 1000 RPM .

3. Membuang supernatan yang terbentuk, kemudian meresuspensikan

endapan (pelet) dengan menambahkan larutan hipotonik hangat KCl

0,075 M, lalu dikocok secara kuat, diresuspensikan homogen dan

diinkubasi pada suhu 37 C dalam waterbath selama 15-30 menit.

4. Melakukan pemusingan kembali pada 1000 RPM selama 10 menit,

lalu membuang supernatan yang terbentuk, dan menambahkan 5ml

larutan fiksasi Carnoys pelan-pelan melalui dinding tabung, kemudian

dikocok.

5. Melakukan pemusingan kembali pada 1000 RPM selama 10 menit,

mengulangi pemberian larutan fiksasi Carnoys sampai didapatkan

presipitat jernih.

6. Melakukan suspensi residu dengan larutan Carnoys secukupnya

sesuai dengan banyaknya pelet yang terbentuk.

7. Meneteskan dan menyebarkan 2 tetes suspensi pada gelas objek pada

lokasi yang berbeda.

8. Mengeringkan gelas objek, lalu mengamati dibawah mikroskop

cahaya.
 67

9. Menghitung jumlah kromosom pada 1 metafase sebagai skrining awal,

lalu penghitungan jumlah kromosom dilakukan sampai 20 metafase.

10. Jika terdapat kesulitan dalam mengamati adanya metafase di bawah

mikroskop cahaya, dilakukan pengecatan dengan giemsa : buffer

phosphate (1:9). Lalu slide dikeringkan lagi, dan diamati kembali

dibawah mikroskop cahaya.

- Pengecatan Trypsin tanpa penghangatan (GTG banding)

1. Mencelupkan slide yang berumur lebih dari 3 hari kedalam larutan

Trypsin 0,1% yang dilarutkan dengan 100 ml PBS pH 6,8 selama 10-

15 detik.

2. Mencuci slide dengan larutan PBS, selanjutnya slide dimasukkan ke

larutan giemsa 10% dalam buffer fosfat (1:9) selama 10-15 menit.

3. Mencuci slide dibawah air mengalir, lalu dikeringkan.

4. Mengamati slide dibawah mikroskop cahaya.

- Analisis

- Analisis dikerjakan oleh dokter ahli sitogenetika dengan menyiapkan

format analisis untuk mencatat koordinat dan jumlah metafase yang

dihitung dan dianalisis.

- Analisis untuk seluruh kasus harus dengan pengecatan G-Banding,

paling sedikit 8 metafase dan penghitungan 20 metafase. Apabila

didapatkan kelainan mosaik, analisis dilakukan pada 100 metafase.

 68

- Prosedur Laporan/Diagnosis

Cara melaporkan bentuk atau konsitusi kromosom adalah mengikuti cara

yang diharuskan oleh International System for Human Cytogenetic

Nomenclature (ICSN). Standar penulisan konsitusi kromosom adalah pertama

kali tulis jumlah kromosom kemudian diikuti koma dan jenis kromosom seks,

diikuti koma lagi dan selanjutnya kelainan struktural (bila terdapat kelainan

struktural). Apabila melibatkan kelainan kromosom pada 2 kromosom maka

ditulis jenis kromosom secara urut nomor yang kecil. Seluruh metafase yang

telah dianalisis, kemudian difoto hitam putih, tiap-tiap kromosom digunting

dan ditempel pada kertas yang sudah disediakan sesuai dengan urut-urutanan

nomernya. Dokter ahli sitogenetika menetukan jenis kariotip serta memberikan

kesimpulan dari hasil pemeriksaan.

 69

Lampiran 3. Hasil Analisis Program Statistik

Case Processing Summary

Cases

Valid Missing Total

N Percent N Percent N Percent

Obat Nyamuk * Isolated
233 93.6% 16 6.4% 249 100.0%
hypospadias
Ayah Perokok * Isolated
233 93.6% 16 6.4% 249 100.0%
hypospadias
Kontrasepsi hormonal *
233 93.6% 16 6.4% 249 100.0%
Isolated hypospadias
Pestisida * Isolated
233 93.6% 16 6.4% 249 100.0%
hypospadias
Penggunaan obat * Isolated
233 93.6% 16 6.4% 249 100.0%
hypospadias
Paritas * Isolated
227 91.2% 22 8.8% 249 100.0%
hypospadias
Usia ibu * Isolated
220 88.4% 29 11.6% 249 100.0%
hypospadias
Berat badan lahir* Isolated
146 58.6% 103 41.4% 249 100.0%
hypospadias

Statistics

Paritas Usia Ibu Berat Badan

Lahir

Valid 235 229 191

N
Missing 14 20 58
Mean 1.58 28.00 2848.99
Median 1.00 27.00 2900.00
Std. Deviation 1.182 6.740 697.358
Minimum 1 16 1000
Maximum 12 50 4500
 70

Penggunaan Obat Nyamuk * Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

Count 9 43 52
Ya % within Isolated
Penggunaan 22.5% 22.3% 22.3%
hypospadias
Obat
Count 31 150 181
Nyamuk
Tidak % within Isolated
77.5% 77.7% 77.7%
hypospadias

Count 40 193 233

Total % within Isolated
100.0% 100.0% 100.0%
hypospadias

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square .001 1 .976
b
Continuity Correction .000 1 1.000
Likelihood Ratio .001 1 .976
Fisher's Exact Test 1.000 .561
Linear-by-Linear Association .001 1 .976
N of Valid Cases 233

a. 0 cells (.0%) have expected count less than 5. The minimum expected count is 8.93.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for penggunaan

1.013 .448 2.290
obat nyamuk (Ya/ Tidak)
For cohort Isolated
1.011 .515 1.985
hypospadias = Berat
For cohort Isolated hypospadias
.998 .867 1.149
= Ringan
N of Valid Cases 233
 71

Ayah Perokok * Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

Count 7 25 32
Ya
Ayah % within Isolated hypospadias 17.5% 13.0% 13.7%
perokok Count 33 168 201
Tidak
% within Isolated hypospadias 82.5% 87.0% 86.3%

Total Count 40 193 233

% within Isolated hypospadias 100.0% 100.0% 100.0%

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square .578 1 .447
b
Continuity Correction .258 1 .611
Likelihood Ratio .547 1 .459
Fisher's Exact Test .452 .296
Linear-by-Linear Association .576 1 .448
N of Valid Cases 233

a. 0 cells (.0%) have expected count less than 5. The minimum expected count is 5.49.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for ayah

1.425 .570 3.568
perokok(Ya/ Tidak)
For cohort Isolated
1.332 .645 2.752
hypospadias = Berat
For cohort Isolated hypospadias
.935 .770 1.134
= Ringan
N of Valid Cases 233
 72

Penggunaan kontrasepsi hormonal* Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

Count 3 23 26
Ya % within Isolated
Penggunaan 7.5% 11.9% 11.2%
hypospadias
Kontrasepsi
Count 37 170 207
Hormonal
Tidak % within Isolated
92.5% 88.1% 88.8%
hypospadias
Count 40 193 233
Total % within Isolated
100.0% 100.0% 100.0%
hypospadias

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square .652 1 .419
b
Continuity Correction .283 1 .595
Likelihood Ratio .712 1 .399
Fisher's Exact Test .584 .311
Linear-by-Linear Association .649 1 .420
N of Valid Cases 233

a. 1 cells (25.0%) have expected count less than 5. The minimum expected count is 4.46.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for penggunaan

.599 .171 2.101
kontrasepsi hormonal(Ya/ Tidak)
For cohort Isolated hypospadias =
.646 .214 1.946
Berat
For cohort Isolated hypospadias =
1.077 .925 1.255
Ringan
N of Valid Cases 233
 73

Paparan Pestisida* Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

Count 8 32 40
Ya % within Isolated
20.0% 16.6% 17.2%
Paparan hypospadias
Pestisida Count 32 161 193
Tidak % within Isolated
80.0% 83.4% 82.8%
hypospadias
Count 40 193 233
Total % within Isolated
100.0% 100.0% 100.0%
hypospadias

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square .272 1 .602
b
Continuity Correction .085 1 .771
Likelihood Ratio .264 1 .607
Fisher's Exact Test .646 .374
Linear-by-Linear
.271 1 .602
Association
N of Valid Cases 233

a. 0 cells (.0%) have expected count less than 5. The minimum expected count is 6.87.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for paparan pestisida(Ya /

1.258 .531 2.980
Tidak)
For cohort Isolated hypospadias = Berat 1.206 .601 2.419
For cohort Isolated hypospadias = Ringan .959 .811 1.134
N of Valid Cases 233
 74

Konsumsi Obat * Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

Count 11 52 63
Ya
Konsumsi % within Isolated hypospadias 27.5% 26.9% 27.0%
Obat Count 29 141 170
Tidak
% within Isolated hypospadias 72.5% 73.1% 73.0%
Count 40 193 233
Total
% within Isolated hypospadias 100.0% 100.0% 100.0%

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square .005 1 .942
b
Continuity Correction .000 1 1.000
Likelihood Ratio .005 1 .943
Fisher's Exact Test 1.000 .541
Linear-by-Linear
.005 1 .943
Association
N of Valid Cases 233

a. 0 cells (.0%) have expected count less than 5. The minimum expected count is 10.82.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for konsumsi obat(Ya /

1.029 .479 2.207
Tidak)
For cohort Isolated hypospadias =
1.024 .545 1.924
Berat
For cohort Isolated hypospadias =
.995 .872 1.136
Ringan
N of Valid Cases 233
 75

Paritas* Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

Count 24 127 151

Primipara
% within Isolated hypospadias 61.5% 67.6% 66.5%
Paritas
Count 15 61 76
Multipara
% within Isolated hypospadias 38.5% 32.4% 33.5%
Count 39 188 227
Total
% within Isolated hypospadias 100.0% 100.0% 100.0%

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square .525 1 .469
b
Continuity Correction .289 1 .591
Likelihood Ratio .516 1 .473
Fisher's Exact Test .463 .292
Linear-by-Linear Association .522 1 .470
N of Valid Cases 227

a. 0 cells (.0%) have expected count less than 5. The minimum expected count is 13.06.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for

.769 .376 1.569
paritas(Primipara / Multipara)
For cohort Isolated
.805 .449 1.443
hypospadias = Berat
For cohort Isolated hypospadias
1.048 .919 1.195
= Ringan
N of Valid Cases 227
 76

Usia ibu * Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

>35 Count 9 20 29
tahun % within Isolated hypospadias 23.1% 11.0% 13.2%
Usia ibu
35 Count 30 161 191
tahun % within Isolated hypospadias 76.9% 89.0% 86.8%
Count 39 181 220
Total
% within Isolated hypospadias 100.0% 100.0% 100.0%

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square 4.056 1 .044
b
Continuity Correction 3.073 1 .080
Likelihood Ratio 3.574 1 .059
Fisher's Exact Test .064 .045
Linear-by-Linear Association 4.037 1 .045
N of Valid Cases 220

a. 0 cells (.0%) have expected count less than 5. The minimum expected count is 5.14.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for usia ibu(>35 tahun / 35

2.415 1.004 5.810
tahun)
For cohort Isolated hypospadias =
1.976 1.048 3.726
Berat
For cohort Isolated hypospadias =
.818 .636 1.052
Ringan
N of Valid Cases 220
 77

Berat badan lahir * Isolated Hypospadias

Crosstab

Isolated hypospadias Total

Berat Ringan

<2500 Count 9 33 42
Berat
gram % within Isolated hypospadias 33.3% 27.7% 28.8%
Badan
2500 Count 18 86 104
Lahir
gram % within Isolated hypospadias 66.7% 72.3% 71.2%
Count 27 119 146
Total
% within Isolated hypospadias 100.0% 100.0% 100.0%

Chi-Square Tests

Value df Asymp. Sig. Exact Sig. Exact Sig.

(2-sided) (2-sided) (1-sided)
a
Pearson Chi-Square .337 1 .562
b
Continuity Correction .119 1 .730
Likelihood Ratio .330 1 .566
Fisher's Exact Test .639 .358
Linear-by-Linear Association .335 1 .563
N of Valid Cases 146

a. 0 cells (.0%) have expected count less than 5. The minimum expected count is 7.77.
b. Computed only for a 2x2 table

Risk Estimate

Value 95% Confidence Interval

Lower Upper

Odds Ratio for berat badan lahir(<2500

1.303 .532 3.189
gram / 2500 gram)
For cohort Isolated hypospadias = Berat 1.238 .605 2.532
For cohort Isolated hypospadias = Ringan .950 .793 1.138
N of Valid Cases 146
 78

Lampiran 4. Biodata Mahasiswa

Identitas
Nama : Khilyatul Mufida
NIM : 22010111120040
TTL : Tuban, 14 April 1993
Jenis Kelamin : Perempuan
Alamat : Perumda IV, 41, Tembalang, Semarang
Nomor Ponsel : +6285731589517
E-mail : [email protected]

Riwayat Pendidikan Formal

1. TK : Tunas Rimba, Tuban, Jawa Timur Lulus Tahun : 1999
2. SD : SDN 1 Sugihan, Tuban, Jawa Timur Lulus Tahun : 2005
3. SMP : SMPN 1 Jatirogo,Tuban, Jawa Timur Lulus Tahun : 2008
4. SMA : SMAN 1 Tuban, Jawa Timur Lulus Tahun : 2011
5. FK UNDIP, Masuk tahun 2011

Keanggotaan Organisasi :

1. Badan Analisis Pengembangan dan Penelitian Kedokteran Indonesia

(BAPIN), ISMKI, Tahun 2014 s.d 2015
2. Asisten Mahasiswa Bagian Anatomi FK Undip Tahun 2013 s.d 2015
3. Kelompok Studi Mahasiswa FK Undip Tahun 2012 s.d 2014
4. Himpunan Mahasiswa FK Undip Tahun 2012 s.d 2014
5. Asisten Mahasiswa Bagian Kimia FK Undip Tahun 2012 s.d 2013
6. Reproduction Health Education Unit FK Undip Tahun 2012 s.d 2013
7. Jaringan Mahasiswa Kesehatan Indonesia Tahun 2012 s.d 2013
8. Rohani Islam FK Undip Tahun 2012 s.d 2013
 79

Pengalaman Penelitian :
1. Prevalensi Infeksi Cacing Cambuk (Trichuris trichiura) dan Cacing Tambang
(Necator americanus dan Ancylostoma duodenale) pada Ibu Hamil di
Kota Semarang, 2013-2014.

Pengalaman Presentasi dan Mengikuti Lomba Karya Ilmiah

1. Pengaruh kombinasi teh hijau dan kulit manggis terhadap profil lipid
serum, foam cell, dan ketebalan aorta : Studi pada Tikus Wistar yang
Diinduksi Aterosklerosis. Departemen Riset BEM FK KM Undip.
[Juara 2 Proposal Penelitian - Fakultas] 2014.
2. Depression Among Indonesian Homosexual Men. FK Universitas
Indonesia. [Semifinalis Research Article - Internasional] 2014
3. Himpunan Mahasiswa Kedokteran Umum Undip. [Finalis 10 Besar
Poster Ilmiah - Nasional]
4. Kombinasi Dietilkarbamazin dan Naringenin dari Kulit Jeruk Purut
(Citrus hystix) melalui Transdermal patch: A Novel Therapy for
Filariasis. Badan Analisis dan Pengembangan Ilmiah Nasional (BAPIN)-
Ikatan Senat Mahasiswa Kedokteran Indonesia (ISMKI) beserta FK
Universitas Padjajaran. [Juara 2 Gagasan Tertulis - Nasional] 2013.
5. Potensi Vaksin Kombinasi Brugia Malayi Abundant Larval Transcript-2
(Bmalt-2) Dan Brugia Malayi Small Heat Shock Protein (Bmhsp) Melalui
Microneedle Patch Terhadap Filariasis Limfatik. Rohani Islam FK
Undip. [Juara 1 Gagasan Tertulis - Nasional] 2013.
6. Restorative Yoga For Breast Cancer Survivors: Physical And
Psychological Benefits, FK Universitas Indonesia. [Semifinalis
Literature Review - Internasional] 2013
7. Kombinasi Melatonin dan Vitamin D sebagai Terapi Target Lupus
Eritematosus Sistemik melalui Modifikasi Sistem Imun. FK Universitas
Sumatera Utara.[Finalis 10 Besar- Nasional] 2012