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"Latar Belakang: Mutasi Al762T/GI764A basal core promoter (BCP) dan Gl896A precore pada genom virus hepatitis B (VHB) berhubungan dengan tingkat keparahan penyakit hati, namun demikian peran mutasi-mutasi tersebut pada perjalanan infeksi hepatitis B kronis masih belum jelas. Olch karena itu, penelitian ini bertujuan untuk mengetahui prevalensi mutasi Al762T/Gl764A dan Gl896A serta hubungannya dengan fase-fase pada perjalanan infeksi hepatitis B kronis. Metodologi: Seratus empat puluh pasien hepatitis B kronis yang dilibatkan dalam pcnelitian ini, belum mendapatkan pengobatan, dan dikelompokkan ke dalam fase immunotolerant (IT), immunoclearance (IC), non/Iow replicative (LR) dan hepatitis "c" negatif (ENH). DNA VHB diperiksa dan diul-cur kadarnya dengan teknik polymerase chain reaction, kemudian disekuensing untuk dianalisis. Hasil: Usia subjek lebih tua pada kelompok ENH dan LR dibandingkan dengan fase lain (p<0.05). Kadar DNA paling tinggi pada fase IC dan paling rendah pada fase LR (p<0.00l), sementara pria mempunyai risiko lebih besar terjadi reaktivasi dengan HBeAg negatif (p<0.05). Mutasi Al 762T/GI764A tidak berbeda bermakna pada semua fase (p=0.56) dan lebih tinggi pada genotipe C dan subtipe adr (p<0.05). Mutasi Gl896A paling tinggi pada Pass LR (p<0.05), dan tidak berbeda bcrmakna pada genotipe dan subtipe VHB. Tidak ada hubungan antara kadar DNA V1-IB dengan mutasi di prccore dan BCP. Kesimpulan: Prevalensi mutasi Gl896A berbeda pada fase hepatitis B kronis di Indonesia, ditemukan lebih sering pada usia lebih tua dan fase lanjut. Mutasi A1762T/Gl764A berkorelasi dengan genotipe dan subtipe VHB, sebaliknya tidak berhubungan dengan fase infeksi. Studi ini mengindikasikan bahwa mutasi BCP tidak berhubungan dcngan serokonversi HBeAg pada perjalanan infeksi hepatitis B kronis.

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Background: Precore Gl896A and basal core promoter (BCP) A1762T/G1764A mutations of hepatitis B virus (HBV) genome have been correlated with severe liver diseases; however, their role in the pathogenesis of chronic hepatitis B (CHB) remains unclear. We assessed the prevalence and association of these mutations in different phases of CHB in Indonesian patients.

Methods: One-hundred and forty CHB patients, not undergoing antiviral therapy, were classified into immune-tolerance (IT), immune-clearance (IC), nonllow- replicative (LR), and hepatitis B e antigen (HBeAg)-negative hepatitis (ENH) phases. HBV DNA was detected and quantified by polymerase chain reaction then analyzed by sequencing.

Results: ENH and LR patients were older than IC or IT patients (p <0.05). HBV DNA levels were highest in IC patients and lowest in LR (p<0.0001). The A1896 pre-core mutants were most prevalent in LR (p<0.00l) and higher in ENH (p<0.00I) than in IT and IC patients, while the Al762T/Gl764A BCP mutants were comparable between all phases. The Al762T/Gl764A BCP mutants were more frequently identified in genotype C than in genotype B (p <0.05), and in subtype adr than in subtypes adw and ayw (p <0.05). The Tl858 mutants were detected in almost all HBV isolates regardless the genotypes (B and C). N0 associations were observed between HBV DNA levels and precore as well as BCP mutations.

Conclusions: The prevalence of precore A1896 mutation differed in phases of CHB in Indonesian patients with preponderance in older ages and later stages. BCP AI762T/Gl764A mutations were associated with HBV genotypes and subtypes, itrespective of infection phases. These findings indicate that BCP mutations could be independent of HBeAg seroconversion in the natural history of chronic HBV infection."

"Karsinoma hepatoseluler (KHS) adalah salah satu kanker dengan laju mortalitas tertinggi di dunia. Kadar serum alfa-fetoprotein (AFP) dapat digunakan sebagai biomarker untuk menegakkan diagnosis dini. Tetapi, perbandingan antara kadar serum AFP dan KHS dengan etiologi infeksi virus dan etiologi non infeksi virus belum diketahui. Mengetahui perbandingan antara kadar serum AFP dan KHS dengan etiologi infeksi virus dan etiologi non infeksi virus. Penelitian potong lintang dilakukan di RSUPN Cipto Mangunkusumo, Jakarta pada Januari-Oktober 2018 dengan melihat data rekam medis dari 287 pasien yang terdiagnosis KHS dalam periode 2013-2017. Nilai median (minimum-maksimum) dari kadar AFP pada pasien KHS dengan etiologi infeksi VHB atau VHC adalah 419 (0.8-400.000). Nilai median (minimum-maksimum) kadar AFP pada pasien KHS dengan etiologi non infeksi VHB-VHC adalah 7.18 (0.6-90.944). Terdapat perbedaan bermakna antara kadar AFP dengan KHS dengan etiologi infeksi VHB atau VHC dan etiologi non infeksi VHB-VHC.

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Hepatocellular carcinoma (HCC) is one of the highest rates of mortality in the world. Serum alpha-fetoprotein (AFP) levels can be used as a biomarker for early diagnosis. However, the comparison between serum AFP and HCC with viral infections etiology and non-viral etiology is unknown. This research aims to determine the comparison between serum AFP and HCC with viral infections etiology and non-viral aetiology. A cross-sectional study conducted in Cipto Mangunkusumo Hospital, Jakarta in January to October 2018 by reviewing 287 medical records of patients diagnosed with HCC from 2013-2017 period of time. The median (minimum-maximum) value of AFP levels in HCC patients with the etiology of HBV or HCV infection is 419 (0.8-400,000). The median value (minimum-maximum) of AFP levels in HCC patients with the etiology of non HBV-HCV infection was 7.18 (0.6-90,944). There were significant differences between AFP levels and KHS with the etiology of HBV or HCV infections and the etiology of non HBV-HCV infections."

"Chronic hepatitis B is still a major health problem in Indonesia. Unfortunately, to date, treatment of chronic HBV (Hepatitis B virus) infection had not shown satisfactory result. Monotherapy with alpha interferon or lamivudine have been widely used as treatment of chronic HBV. However, treatment response to Alpha interferon in Asian people was not satisfactory (15% - 20%), while monotherapy with lamivudine was not sufficient to eradicate HBV in chronically infected patients and commonly induce drug resistance. The occurrence of chronic hepatitis B resistant to lamivudine had encouraged development of newer agents such as adefovir, entecavir, emtricitabine and nucleoside analog. New therapeutic strategy using combination therapy should be considered if there is no sufficient response to monotherapy"

"ABSTRAKInfeksi oleh virus hepatitis B (VHB) dapat menjadi infeksi akut yang berakhir dengan resolusi infeksi ataupun berlanjut menjadi infeksi kronis. Resolusi infeksi dalam infeksi VHB ditandai dengan hilangnya hepatitis B surface antigen (HBsAg) dan keberadaan antibodi terhadap HBsAg (anti-HBs). Kemampuan sel B dalam mensintesis anti-HBs dipengaruhi oleh sel T helper 1 (Th1) ataupun T helper 2 (Th2). Sekresi sitokin yang terkoordinasi dari Th1 ataupun Th2 sangat dibutuhkan mengingat sitokin yang dihasilkan oleh kedua sel T helper (Th) tersebut memiliki peranan yang berbeda dan dapat bekerja secara antagonis. Penelitian ini bertujuan untuk mengetahui kemampuan pasien hepatitis B kronis dalam mensintesis anti-HBs dan membandingkan pola sintesis sitokin IL-10, IFN-gamma dan IL-2 dari pasien hepatitis B kronis dengan pasien yang mengalami resolusi infeksi. Pada penelitian ini sel mononuklear darah tepi manusia (SMDT) diambil dari 10 subjek pasien hepatitis B kronis, 10 subjek pasien yang mengalami resolusi infeksi dari hepatitis B dan 10 subjek individu sehat yang berhasil divaksinasi. SMDT dikultur dengan stimulan HBsAg rekombinan (rHBsAg) atau fitohemaglutinin (PHA) untuk sintesis sitokin dan pokeweed mitogen (PWM) untuk sintesis anti-HBs secara in vitro. Hasil dari penelitian ini ditemukan produksi anti-HBs secara in vitro dari 70% individu sehat yang berhasil divaksinasi dan 40% pasien hepatitis B yang mengalami resolusi infeksi, sementara pada pasien hepatitis B kronis tidak ditemukan hal tersebut. Pola sitokin IL-10, IFN-gamma dan IL-2 antara pasien hepatitis B yang mengalami resolusi infeksi dan pasien hepatitis B kronis tidak memiliki perbedaan yang bermakna. Akan tetapi, respons IFN-gamma terhadap rHBsAg pada pasien hepatitis B yang mengalami resolusi infeksi cenderung lebih kuat. Korelasi antara sitokin IL-10, IFN-gamma dan IL-2 dengan produksi anti-HBs secara in vitro tidak ditemukan pada kedua kelompok tersebut, sementara korelasi IL-2 dengan sintesis anti-HBs in vitro ditemukan pada individu yang divaksinasi. Penelitian ini menunjukkan SMDT dari pasien hepatitis B kronis tidak dapat mensintesis anti-HBs secara in vitro dan pada pasien tersebut tidak memiliki perbedaan pola sintesis sitokin IL-10, IFN-gamma dan IL-2 jika dibandingkan dengan pasien hepatitis B yang mengalami resolusi infeksi.

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ABSTRACTHepatitis B virus (HBV) infection can lead to acute self-limited infection or lead to chronic hepatitis B infection. Resolution of infection is marked by seroconversion of hepatitis B surface antigen (HBsAg) to antibody to HBsAg (anti-HBs) which also give protection to HBV reinfection. Anti-HBs is produced by B cells as response to HBsAg. B cells response to HBsAg is affected by cytokines from T helper 1 (Th1) and T helper 2 (Th2) cells. Coordinated cytokines secreted by Th1 or Th2 cells is necessary due to the fact that they could work antagonistically. Th1 cytokines, such as IFN-gamma, are known to induce cellular immune responses, while Th2 cytokines, such as IL-4, -5 and -10, are known to induce humoral immune responses. This study aimed to investigate the capability of chronic hepatitis B patients (CHB) to synthesize anti-HBs in vitro and to compare IL-10, IFN-gamma and IL-2 levels between CHB patientis with resolved hepatitis B (RHB) patients. In this study, peripheral blood mononuclear cells (PBMCs) were taken from 10 CHB patients, 10 RHB patients and 10 healthy hepatitis B vaccinated individuals. PBMCs were cultured in presence of recombinant HBsAg (rHBsAg) and PHA to cytokines synthesis and pokeweed mitogen (PWM) to anti-HBs synthesis in vitro. As results, synthesis of anti-HBs in vitro were found in PBMCs from 70% of healthy hepatitis B vaccinated individuals and 40% of RHB patients, while PBMCs from CHB patients could not. No significant differences were found in IL-10, IFN-gamma and IL-2 cytokine levels between CHB patients and RHB patients, although IFN-gamma responses to rHBsAg had a tendency to be stronger in RHB patients. Correlation between IL-10, IFN-gamma and IL-2 cytokine levels and anti-HBs synthesis in vitro was not found in CHB patients and RHB patients. Meanwhile, IL-2 and anti-HBs synthesis in vitro were correlated in healthy hepatitis B vaccinated individuals. In conclusion, this study showed that PBMCs from CHB patients were not capable in synthesizing anti-HBs in vitro and had no differences in IL-10, IFN-gamma and IL-2 cytokine levels with RHB patients."

"Amplifikasi DNA virus hepatitis B (VHB) dari sampel plasma sulit dilakukan apabila kadar DNA VHB <10.000 kopi/ml. Sehingga, langkah awal ekstraksi menjadi penting bagi keberhasilan amplifikasi dan sekuensing. Peningkatan konsentrasi DNA dalam jumlah yang cukup dan berkualitas baik memerlukan metode isolasi yang optimal. Tujuan penelitian ini adalah mendapatkan teknik isolasi DNA yang optimal untuk menghasilkan DNA VHB dalam jumlah yang cukup sehingga dapat digunakan untuk mendeteksi perubahan genetik gen polimerase VHB terkait resistensi obat antivirus pada kasus hepatitis B kronik. Optimasi prosedur penanganan sampel untuk mengekstraksi partikel DNA VHB dilakukan dengan mengisolasi sampel plasma dengan kadar virus 105 kopi/ml menggunakan delapan perlakuan yang berbeda yaitu 1 ml dan 200 µL plasma disentrifugasi 4000xg selama 20 menit pada suhu 25°C (P1 dan P2), 1 ml dan 200 µL plasma disentrifugasi 16000xg selama 1 jam pada suhu 4°C (P3 dan P4), 1 ml plasma disentrifugasi 21000xg selama 1 jam pada suhu 4°C (P5), 1 ml plasma dalam PEG6000 20% diinkubasi semalaman pada 2-8°C kemudian disentrifugasi 21000xg selama 1 jam pada suhu 4°C (P6), 1 ml plasma dalam PEG6000 20% dan NaCl 0.5M diinkubasi semalaman pada 2-8oC kemudian disentrifugasi 21000xg selama 1 jam pada suhu 4°C (P7), dan 200 µL sampel plasma tanpa penambahan perlakuan (P8). Kadar virus diukur menggunakan kuantitatif realtime PCR, dan hasil dari setiap perlakuan sampel dibandingkan dengan kontrol (P8). Uji simulasi dilakukan pada sampel plasma dengan kadar virus 105; 104; 103 dan 102 kopi/ml yang diberikan perlakuan 3 (P3) yang dipilih berdasarkan hasil sebelumnya, DNA kemudian disekuensing untuk dianalisis mutasi resistensinya terhadap obat antivirus dan hasilnya dibandingkan dengan sampel yang tanpa diberikan perlakuan. Hasil menunjukkan bahwa terjadi peningkatan konsentrasi DNA pada rerata sampel yang diisolasi menggunakan P3 jika dibandingkan dengan kontrol (P8). Sedangkan hasil sekuensing untuk analisis mutasi pada gen polimerase terkait resistensi terhadap obat antivirus dapat dilakukan pada sampel dengan kadar virus 105; 104; 103 kopi/ml baik yang diberikan perlakuan maupun tanpa perlakuan. Namun, sampel dengan kadar virus 102 kopi/ml hanya dapat dianalisis mutasi resistensinya pada sampel yang ditambahkan perlakuan 3 (P3). Kesimpulan: 1 ml plasma yang disentrifugasi 16000xg selama 1 jam pada suhu 4°C (perlakuan 3) merupakan metode isolasi DNA yang mampu meningkatkan perolehan DNA secara optimal dari sampel plasma penderita hepatitis B kronik, sehingga analisis resistensi terhadap obat antivirus menggunakan teknik in-house assay dapat di lakukan pada sampel dengan kadar virus <10.000 kopi DNA/ml.

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The amplification of hepatitis B virus DNA from plasma samples is difficult when HBV DNA levels <10.000 copies/ml. Thus, the initial step of extraction become crucial for the success of amplification and sequencing. Increasing concentrations of DNA in sufficient quantity and good quality need an optimal isolation method. The aim of this study is to obtain an optimal DNA isolation technique and generate HBV DNA in sufficient quantities, so that it can be used to detect genetic changes of HBV polymerase gene related to drugs resistance on chronic hepatitis B. The optimization of sample handling procedure for extracting HBV DNA particles were performed by isolating plasma samples with viral load 105 copies/ml using eight different kinds of treatment are 1 ml and 200 µL of plasma was centrifuged at 4000×g for 20 minutes at 25°C (P1 and P2); 1 ml and 200 µL of plasma centrifuged at 16000×g for 1 hour at 4°C (P3 and P4); 1 ml of plasma centrifuged at 21000×g for 1 hour at 4°C (P5); 1 ml of plasma in 20% PEG6000 were incubated overnight at 2-8°C then centrifuged at 21000xg for 1 hour at 4°C (P6), 1 ml of plasma in 20% PEG6000 and 0.5M NaCl were incubated overnight at 2-8oC then centrifuged at 21000xg for 1 hour at 4°C (P7), and 200 µL of plasma without treatment (P8). Hepatitis B virus level were measured using quantitative real-time PCR, and the results of each treatment compared to the control samples (P8). Simulation test performed on plasma samples with grading of virus level (i.e 105; 104; 103 and 102 copies/ml) were given treatment 3 (P3) were selected based on previous results, the DNA was sequenced and then analyzed for drugs resistance and the results were compared with samples without treatment.

Result showed that an increasing in average concentration of DNA samples was isolated using the P3 when compared with controls (P8). While the results of sequencing for analysis of mutations in polymerase gene associated with drugs resistance can be performed on samples with virus level 105; 104; 103 copies/ml were given either treatment or no treatment. However, samples with virus level 102 copies/ml can be analyzed only

on treatment samples.

Conclusion: 1 ml plasma were centrifuged at 16000xg for 1 h at 4°C (treatment 3) is a DNA isolation method that can improve the recovery of optimal DNA from plasma samples of patients with chronic hepatitis B, so that the analysis of drug resistance using in-house assay techniques can be done on samples with virus levels <10,000 copies/ml."

"ABSTRAK<

Karsinoma hepatoseluler (KHS) merupakan karsinoma primer tersering pada sel hati. Sebagian besar KHS disebabkan oleh virus hepatitis B (VHB) dan virus hepatitis C (VHC) yang memiliki patogenesis yang berbeda dalam menyebabkan KHS. Alfa-fetoprotein (AFP) sebagai penanda tumor pada KHS dan dipengaruhi oleh berbagai faktor, salah satunya status infeksi. Berbagai penelitian sudah dilakukan untuk mengetahui pengaruh pengaruh jenis virus penyebab KHS dengan kadar AFP namun hasilnya sangat beragam. Berdasarkan hal tersebut dan ditambah dengan belum adanya penelitian serupa yang menggunakan data pasien di Indonesia maka penelitian ini bertujuan untuk membandingkan kadar AFP pada pasien KHS terkait infeksi VHB terhadap VHC. Penelitian ini dilakukan dengan desain studi potong lintang menggunakan 199 data AFP pasien KHS yang terdiri dari 129 kasus KHS terkait VHB dan 70 kasus KHS terkait VHC. Dari penelitian ini didapatkan sebanyak 97% dan 87.3% pasien KHS terkait VHC dan VHB mengalami peningkatan kadar AFP secara berurutan. Nilai median kadar AFP pada pasien KHS terkait VHB adalah 419 IU/mL sedangkan pada pasien KHS terkait VHC sebesar 400 IU/mL. Perbedaan nilai tersebut memiliki nilai p = 0.97 dalam uji Mann-Whitney U sehingga disimpulan tidak ada perbedaan bermakna pada rerata kadar AFP antara pasien KHS terkait VHB dibanding dengan VHC.

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ABSTRACT

Hepatocellular carcinoma (HCC) is the most primary common carcinoma in liver cells. Most HCC are caused by the hepatitis B virus and hepatitis C that have different pathogenesis in causing carcinoma. Alpha-fetoprotein as tumor marker in HCC is influenced by various factors, one of which is infection status. Various studies have been carried out to determine the influence of the types of viruses causing HCC with AFP levels but the results are very diverse. Based on this and coupled with the absence of similar studies using patient data in Indonesia, this study aims to compare AFP levels in HCC patients related to HBV and HCV. Using cross-sectional design, this study included 199 data of AFP in patient with HCC comprises of 129 cases of HCC related to HBV and 70 cases of HCC related to HCV. From this study, it was found that 97% and 87.3% of HCC patients related to HCV and HBV experienced an increase in AFP levels consecutively. The median value of AFP levels in HBV-related HCC patients was 419 IU / mL while in HCV-related HCC patients was 400 IU / mL. The difference in value has a p value = 0.97 in the Mann-Whitney U test thus it is concluded that there is no significant difference in AFP levels between HBV-related HCC patients compared with HCV-related HCC.

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"[Latar belakang. Infeksi virus hepatitis B (VHB) di Indonesia masih tinggi dengan rerata prevalensi 9,4%. Tingginya prevalensi HB ini terkait dengan terjadinya infeksi VHB pada masa dini kehidupan, terutama melalui transmisi vertikal. Di Indonesia proporsi transmisi vertikal 45,9% dan 5,2% ibu hamil HBsAgnya positif. Cara paling efektif mengontrol infeksi VHB adalah dengan imunisasi, namun terdapat perbedaan seroproteksi titer anti-HBs pada usia lebih dari 10 tahun di berbagai tempat. Selain itu terdapat faktor-faktor yang dapat memengaruhi titer anti-HBs, namun penelitian ini masih jarang dan belum pernah dilakukan di Manado.Tujuan. Mengetahui seroproteksi titer anti-HBs dan faktor-faktor yang dapat memengaruhi titer anti- HBs tersebut.Metode. Penelitian analitik observational dengan desain potong lintang. Penelitian dilakukan dengan stratified random sampling pada usia 10-15 tahun di Kecamatan Tuminting, Kota Manado sejak Oktober sampai November 2014. Data dianalisis dengan SPSS 22.Hasil. Dari 48 sekolah terpilih 10 sekolah dengan 105 anak sebagai subyek penelitian, namun hanya 23 anak yang mempunyai seroprotektif (21,9%). Sebanyak 76 (72,4%) subyek adalah perempuan, 78 (74,3%) subyek berstatus gizi baik dan 98 (93,3%) subyek memiliki berat badan lahir ≥ 2.500 gram. Dari buku imunisasi didapatkan 26 (24,8%) subyek dengan vaksinasi HB-1 ≤ 7 hari dan 45 (42,9%) subyek dengan jarak HB-2 dan HB-3 ≥ 2 bulan. Didapatkan 86 (81,9%) ibu subyek berusia 20-35 tahun, 64 (60,9%) ibu subyek berpendidikan SMA dan 79 (75,2%) orangtua subyek berpenghasilan ≥ 2 juta per bulan. Analisis multivariat didapatkan faktor pemberian HB-1 < 7 hari atau ≥ 7 hari (p=0,02) dan jarak pemberian HB-2 dengan HB-3< 2 bulan atau ≥ 2 bulan (p<0,001) berperan terhadap seroproteksi HB pada anak.Simpulan. Penelitian ini mendapatkan angka seroproteksi HB yang rendah (21,9%) serta faktor pemberian HB-1 ≤ 7 hari atau > 7 hari dan jarak pemberian HB-2 dengan HB-3 < 2 bulan atau ≥ 2 bulan berperan terhadap seroproteksi HB pada anak usia 10-15 tahun.;Background. Hepatitis B viral (HBV) infection in Indonesia is still high with average prevalence of 9.4%. The high prevalence of hepatitis B (HB) is related to the occurence of HBV infection during the early life, especially through vertical transmission. In Indonesia proportion of vertical transmission 45.9% and 5.2% pregnant women have HBsAg positive. The most effective way to control HBV infection is with immunization HB, but there is differential in anti-HBs seroprotection titer at the age more than ten years in many locations. In addition there are factors that can affect anti-HBs titer, but these studies are rare and have ever been done in Manado.Objective. Knowing anti-HBs seroprotection titer and factors that can affect the anti-HBs titer.Method. Analitic observational study with cross sectional design. Research was done with stratified random sampling in children age 10-15 years old at Tuminting district, Manado city since October until November 2014. Analise data with SPSS 22.Results. From 48 schools, selected 10 schools with 105 children as subject of research, but only 23 (21.9%) children who were having seroprotective (21,9%). A total of 76 (72.4%) subjects were female, 78 (74.3%) subjects with good nutrition status and 98 (93.3%) subjects had ≥2,500 grams birth weight. From the immunization record book 26 (24.8%) subjects were obtained with HB-1 vaccination done at ≤7 days of age and 45 (42.9%) subjects with the distance between HB-2 and HB-3 were ≥2 months. Mother’s age was found 86 (81.9%) were 20-35 years old, 64 (60.9%) mothers’s education were high school graduated and 79 (75.2%) parents subjects had income ≥2 million per month. From multivariate analysis obtained that administration of HB-1 ≤7 days or >7 days (p=0.02) and distance between administration of HB-2 and HB-3 <2 months or ≥2 months (p<0.001) had important role in HB seroprotection in children.Conclusion. This study obtained a number of low HB seroptotection (21.9%) as well as administration of HB-1 ≤7 days or >7 days and distance between administration of HB-2 and HB-3 <2 months or ≥2 months had important role in HB seroprotection in children age 10-15 years old., Background. Hepatitis B viral (HBV) infection in Indonesia is still high with average prevalence of 9.4%. The high prevalence of hepatitis B (HB) is related to the occurence of HBV infection during the early life, especially through vertical transmission. In Indonesia proportion of vertical transmission 45.9% and 5.2% pregnant women have HBsAg positive. The most effective way to control HBV infection is with immunization HB, but there is differential in anti-HBs seroprotection titer at the age more than ten years in many locations. In addition there are factors that can affect anti-HBs titer, but these studies are rare and have ever been done in Manado.Objective. Knowing anti-HBs seroprotection titer and factors that can affect the anti-HBs titer.Method. Analitic observational study with cross sectional design. Research was done with stratified random sampling in children age 10-15 years old at Tuminting district, Manado city since October until November 2014. Analise data with SPSS 22.Results. From 48 schools, selected 10 schools with 105 children as subject of research, but only 23 (21.9%) children who were having seroprotective (21,9%). A total of 76 (72.4%) subjects were female, 78 (74.3%) subjects with good nutrition status and 98 (93.3%) subjects had ≥2,500 grams birth weight. From the immunization record book 26 (24.8%) subjects were obtained with HB-1 vaccination done at ≤7 days of age and 45 (42.9%) subjects with the distance between HB-2 and HB-3 were ≥2 months. Mother’s age was found 86 (81.9%) were 20-35 years old, 64 (60.9%) mothers’s education were high school graduated and 79 (75.2%) parents subjects had income ≥2 million per month. From multivariate analysis obtained that administration of HB-1 ≤7 days or >7 days (p=0.02) and distance between administration of HB-2 and HB-3 <2 months or ≥2 months (p<0.001) had important role in HB seroprotection in children.Conclusion. This study obtained a number of low HB seroptotection (21.9%) as well as administration of HB-1 ≤7 days or >7 days and distance between administration of HB-2 and HB-3 <2 months or ≥2 months had important role in HB seroprotection in children age 10-15 years old.]"