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Abstrak :
[ABSTRAK
Latar belakang. Hepatitis B merupakan salah satu masalah kesehatan yang serius, diperkirakan lebih dari 2 milyar orang didunia telah terinfeksi virus hepatitis B (VHB). Dari jumlah ini kira-kira 360 juta orang mengalami infeksi khronis. Kematian terutama disebabkan karena sirosis hepatis dan karsinoma hepatoseluler. Salah satu upaya pencegahan penularan infeksi VHB adalah uji saring darah donor terhadap hepatitis B surface antigen (HBsAg) yang merupakan pemeriksaan skrinning yang dilakukan oleh Unit Transfusi Darah (UTD) di negara berkembang seperti Indonesia. Banyak peneliti membuktikan bahwa darah HBsAg negatif masih berpotensi menularkan infeksi VHB. Untuk itu meningkatkan keamanan darah, beberapa negara menambahkan parameter pemeriksaan antibodi terhadap hepatits B core (anti-HBc) sebagai petanda paparan terhadap infeksi VHB dan pemeriksaan antibodi terhadap hepatitis B surface (anti-HBs) sebagai tanda respon imun terhadap infeksi VHB. Dengan berkembangnya teknologi biologi molekuler, masa jendela infeksi VHB dengan seronegatif dapat di ketahui lebih cepat melalui deteksi DNA VHB dengan metode Nucleic Acid Test (NAT) multipleks yang dilanjutkan dengan NAT discriminatory. Beberapa penelitian terdahulu menunjukkan, didapatnya DNA VHB pada spesimen darah donor yang seronegatif dengan metoda NAT. Darah dengan HBsAg negatif, DNA VHB positif dengan atau tanpa anti-HBc dan atau anti-HBs merupakan darah asal donor dengan Hepatitis B Occult (HBO). Prevalensi donor dengan HBO di Indonesia berkisar antara 8- 10%. Metodologi. Penelitian ini menggunakan desain potong lintang (cross sectional) yang dilakukan di UTDP dan Lembaga Biologi Molekul Eijkman, dengan jumlah sampel 4.973 asal subyek donor darah dari 4 UTD daerah DKI, Kota Tanggerang,kota Depok dan Kabupaten Tanggerang. Terhadap sampel penelitian dilakukan pemeriksaan serologis HBsAg,anti-HBc,anti-HBs, NAT, dan PCR kuantitatif dan kualitatif, selanjutnya pada sampel yang HBsAg negatif, NAT positif dan didapatkan hasil PCR kuantitatif positif dilakukan pemeriksaan lanjutan sequencing asam amino untuk mengetahui ada tidaknya mutan HBsAg penyebab lolosnya deteksi serologi HBsAg oleh reagensia HBsAg yang digunakan untuk uji saring darah donor. Hasil. Didapatkan hanya 20 subjek (0,40%) mempunyai hasil HBsAg negatif dan NAT positif multipleks, dan hanya 16 subyek (80%) HBsAg negatif dan NAT discrimenatory positif. Lebih lanjut hasil pemeriksaan anti-HBc negatif dan anti- HBs positif/negatif didapatkan hanya1 subyek (6,25%), anti-HBc positif dan anti- Hbs negatif didapatkan 9 subyek (56,25%), hasil pemerksaan anti-Hbc dan anti- HBs positif 5 subyek (31,25%). Lebih lanjut dilakukan pemeriksaan PCR kualitatif dan didapatkan 3 subyek (18,75%) tidak terdeteksi, , 6 (37,5%) subyek menunjukkan hasil viral load yang low detection (dibawah sensitivitas alat), dan 7 subyek (43,75%) menunjukkan hasil viral load dapat di ketahui. Pada pemeriksaan PCR kualitatif dan sequencing didapatkan 2 subyek (28,57%) ditemukan mutasi pada gen S pada posisi 143 dimana terjadi subsitusi asam amino T143M.

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ABSTRACT
Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection. And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg. Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donors Result. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection. And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg. Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donors Result. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection. And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg. Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donors Result. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection. And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg. Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donors Result. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection. And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg. Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donors Result. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution., Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection. And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg. Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donors Result. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.]

Abstrak :
ABSTRAK
Anti-HCV menjadi marker serologi utama yang digunakan untuk uji saring hepatitis C pada donor darah di Indonesia. Selain serologi anti-HCV, untuk lebih meningkatkan kemananan darah, Unit Transfusi Darah UTD DKI Jakarta juga menerapkan pemeriksaan Nucleic Acid Test NAT . Pemeriksaan anti-HCV tidak dapat membedakan antara infeksi aktif dan infeksi yang telah sembuh. Darah akan dianggap terifeksi HCV apabila salah satu dari pemeriksaan serologi atau molekuler positif, begitupula dengan darah donor dengan hasil pemeriksaan anti-HCV grayzone dan NAT negatif. Diperlukan kepastian atas berisiko tidaknya darah tersebut dalam menularkan infeksi HCV, mengingat kebutuhan darah di sebagian besar provinsi di Indonesia masih belum memenuhi target. Sehingga dibutuhkan uji molekuler lain untuk dijadikan pembanding dengan hasil NAT. Interpretasi hasil anti-HCV dilakukan berdasarkan rasio S/CO, yang dapat dijadikan prediksi status viremia donor, sehingga perlu dilakukan analisis hubungan antara S/CO dengan hasil pengujian molekuler dan HCV Ag-Ab. Nilai prediksi viremia diharapkan dapat menjadi alternatif bagi UTD yang belum mampu menerapkan NAT. Kemudian dipilih 93 sampel dengan kriteria anti-HCV positif dan NAT positif; anti-HCV positif dan NAT negatif serta anti-HCV grayzone dan NAT negatif untuk diuji dengan nested PCR kualitatif dan HCV Ag-Ab. Berdasarkan perbandingan hasil pengujian NAT dan nested PCR diperoleh nilai sensitivitas NAT sebesar 90, 63 , dengan Spesifisitasnya 96,71 . Dari hasil analisis chi-square diperoleh hubungan yang bermakna antara nilai S/CO anti-HCV dengan hasil pengujian NAT, nested PCR kualitatif dan HCV Ag-Ab P5 dapat dijadikan prediksi adanya infeksi aktif pada donor.

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ABSTRACT
Anti HCV is the main serological marker for hepatitis C screening in blood donors in Indonesia. Besides anti HCV, UTD DKI Jakarta also implementing Nucleic Acid Test NAT to improve blood transfusion safety. Anti HCV assay can not distinguish between active infection and cured infection. Blood will be considered HCV infected if either from a positive serologic or molecular test, including blood with anti HCV grayzone and NAT negative. There is a requirement to ensure the risk status of blood with anti HCV grayzone and NAT negative, because the supply of blood in most provinces in Indonesia still insufficient. So, it takes another molecular test to compare with NAT result. Interpretation of anti HCV results was calculating by S CO ratio, which could be a predictor of viremia status. It is necessary to analyze the correlation between S CO with molecular test and HCV Ag Ab results.Viremia prediction value is expected to be an alternative for UTDs who have not been able to apply NAT. There are 93 samples collected then tested with NAT and anti HCV. Sample with concondantly positive anti HCV and NAT anti HCV positive and NAT negative and anti HCV grayzone and NAT negative. These samples then tested with nested PCR and HCV Ag Ab. Based on comparison of NAT and nested PCR, obtained NAT sensitivity value of 90, 63 , with Specificity 96.71 . The result of chi square analysis shows a significant correlation between S CO anti HCV with NAT, qualitative nested PCR and HCV Ag Ab P 5 can be used as predictors of active infection in donors.