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"ABSTRAKHepatitis virus adalah lesi inflamasi hati yang difus, hampir selalu disertai kelainan klinis dan biokimia, dan disebabkan oleh infeksi virus. Hepatitis virus akut di Indonesia merupakan penyakitendemis, hampir sepanjang tahun di rumah-rumah sakit kita dapat menemukan penderita penyakit ini. Dari ketiga jenis hepatitis virus utamatersebut diatas, hepatitis virus A tidak pernah menjadi kronis sedang-kan hepatitis B dan NANB dapat menjadi kronis.Tujuan penelitian ini adalah (1). memperoleh data persentase hepatitis virus A akut, B Akut, dan yang mungkin NANB di RS Persahabatan. (2). Memperoleh data gambaran klinis dan pola enzim hepatitis virus A,B, dan NANB akut. (3). Memperoleh data banyaknya penderita hepatitis virus akut yangdirawat di RS Persahabatan setiap bulan. (4). Memperoleh data kronisitasdari masing-masing hepatitis virus akut tersebut.

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"Seperti negara-negara di dunia, di Indonesia penyakit infeksi hepatitis A Virus (HAY) hingga saat ini masih merupakan masalah kesehatan yang besar. Tingginya angka endemisitas dan maiden HAV mempunyai korelasi dengan tingkat higiene dan kondisi sanitasi, disamping perilaku hidup bersih dan sehat (PHBS) masyarakat itu sendiri. Secara umum penularan hepatitis A yang paling dominan adalah secara faecal/ oral melalui makanan dan minuman yang terkontaminasi oleh tinja manusia yang mengandung HAV. Di Kabupaten Kebumen, KLB infeksi HAV yang terjadi pada tahun 2001 menunjukkan kemungkinan masih rendahnya kondisi sanitasi dan PHBS dari masyarakat. Disamping itu masih rendahnya cakupan SAB sebesar 33.3% dan SAGA 16,7% merupakan kontribusi yang patut dipertimbangkan.Tujuan penelitian ini untuk mengetahui faktor-faktor yang berhubungan dengan kejadian infeksi HAV.Rancangan penelitian menggunakan desain kasus kontrol tidak berpadanan, dengan jumlah sampel sebanyak 154 orang. Data sekunder berupa data kasus dan kontrol diperoleh dari hasil pemeriksaan serologic oleh Tim Terpadu (Subdit Surveilans, US NAMRU-2 dan DKK Kebumen) yang berasal dari 2 wilayah puskesmas tempat terjadinya KLB infeksi HAV. Sedangkan data primer dikumpulkan dengan mengunjungi responden untuk melakukan wawancara dan pengamatan menggunakan kuisioner disamping pengambilan sampel air, untuk selanjutnya dianalisis menggunakan program komputer di Laboratorium Komputer Fakultas Kesehatan Masyarakat Universitas Indonesia.Hasil analisis bivariat didapat bahwa terdapat hubungan bermakna antara beberapa faktor risiko yaitu: kualitas bakteriologis air, tempat/sarana b.a.b., cuci tangan setelah b.a.b. dan kebiasaan makan jajan (masing-masing mempunyai nilai pO,05). Hasil analisis multivariat (uji regresi logistik) menunjukkan bahwa faktor dominan yang berhubungan dengan kejadian infeksi HAV adalah: jenis sarana b.a.b., kebiasaan cuci tangan setelah b.a.b. dan makan jajan. Pada penelitian ini tidak ditemukan adanya interaksi antara ketiga variabel tersebut.

Kesimpulan dari penelitian iai adalah bahwa orang yang melakukan b.a.b. di JAGA yang tidak memenuhi syarat, mempunyai kebiasaan tidak mencuci tangan setelah b.a.b.(tidak higienis) dan sering makan jajan berisiko lebih besar terserang infeksi HAV. Disarankan kepada Dinas Kesehatan untuk lebih meningkatkan kegiatan inspeksi sanitasi SAS, secara jangka panjang meningkatkan cakupan SAB & JAGA, melakukan kaponisasi sumur penduduk setelah terjadinya KLB, pemberdayaan masyarakat dan penyuluhan khususnya PHBS dan lingkungan yang saniter. Kepada masyarakat termasuk pengelola usaha makanan jajanan untuk selalu menjaga higiene perorangan dan meningkatkan higiene sanitasi agar kasus serupa tidak terjadi di masa mendatang. Penelitian lanjutan perlu dilakukan untuk mengetahui kekuatan hubungan sebab akibat, disamping memperhatikan variabel-variabel lain yang terkait.

Daftar bacaan: 47 ( 1985-2001)

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Related Factors With Infection Hepatitis A Virus (HAV) Occurence in Regency Kebumen, Year 2001 (Case Control Study of Outbreak Hepatitis)Like nations in the world in Indonesia infection hepatitis A virus (HAV) disease till in this time still represent big health problem. Number endemisitas incident HAV and height have correlation with level of hygiene sanitation and beside the clean life behavior and make healthy (PHBS) society itself In general most dominant infection HAV is by faecal oral of through food and beverage which contamination by faeces of human being containing HAV. That happened outbreak infection HAV in regency Kebumen the year 2001 showing the possibility of still lower sanitation and PHBS from society. Lower of coverage of water supply (SAB) equal to 33,3% and family latrine (SAGA) 16,7% representing proper contribution considered. Aims of this research to know epidemiological description and related factors with infection HAV occurrence.

Design of research use not matched case control, with amount of sample as much 154 people. Secondary data of case and control obtained from result inspection of serologic by Inwrought Team (Subdit Surveillance, US NAMRU-2 and DKK Kebumen) coming from 2 region of Public Health Service of place the happening of outbreak infection HAV. While of primary data collected visitedly is respondent to conduct interview and perception use questioner beside take of sample water to know quality of bacteriological Analysis to use computer program in Computer Laboratory of Public Health Faculty University of Indonesia.

Result of bivariate analysis got that there are relation have a meaning of between some risk factor that is quality of water bacteriological, place of excrement (b.a.b), clean hand habit after b.a.b, eat junk food habit and age (each having p value <0,05) with infection HAV occurrence. While other dissimilar risk factor that type of SAS, clean hand habit of before eating and the gender do not related significant (each having p value >0,05). Result of analysis multivariat (Logistic Regression test)) indicating that dominant factors which deal with infection HAV occurrence is: place of b.a.b. type, habit clean hand after b.a.b. and eat junk food habit. At this research is not found an interaction existence of among third the variable.

Conclusion from this research is that one who do b.a.b in JAGA of ineligible, having habit do not clean hand after b.a.b (is not hygienic) and often eat something of the home have risk more attacked by infection HAV. Suggested to Public Health Service to more improve early warning system (SKID), empowerment and the counseling especially of PHBS and healthy environment. To society include the junk food handler always take care of personal hygiene and sanitation in order the similar case is not happened in the future. Continuation research require to be conducted to know causality strength, while considered other variable that related.

Library list: 47 (1985-2001)"

"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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"Hepatitis A merupakan penyakit tertua yang penularannya melalui jalur faecal-oral. Kejadian hepatitis A di RS. Awal Bros Bekasi cenderung meningkat setiap tahunnya. Tujuan penelitian ini adalah mengetahui gambaran epidemiologi hepatitis A di RS. Awal Bros Bekasi Januari 2009 - November 2012. Desain penelitian ini seri kasus. Pemilihan sampel dengan purposive sampling. Berdasarkan hasil penelitian didapatkan penderita hepatitis A berjumlah 93 orang, rata-rata umur penderita 24,44 tahun, didominasi oleh jenis kelamin laki-laki. Lama rawat penderita tergolong pendek. Kasus meningkat pada kuartal keempat tahun 2011 dan kuartal pertama tahun 2012. Berdasarkan data tersebut, diharapkan RS. Awal Bros Bekasi dapat lebih mengoptimalkan pelayanan terhadap penderita hepatitis A.

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Hepatitis A is the oldest disease with faecal-oral trasmission. The inicidence of Hepatitis A in Awal Bros Hospital, Bekasi tend to increase every year. The purpose of this research was to investigate epidemiology description of hepatitis A in Awal Bros Hospital, Bekasi January 2009 - November 2012. This research used case series design. The results show that from 93 respondent, the average age of 24,44 years old. The majority gender is the male, and average length of stay of the respondent is short. Cases increased in the fourth quarter of 2011 and first quarter of 2012. From this research, it was concluded that Awal Bros Hospital, Bekasi can be better for the service of people with hepatitis A."

"Latar Belakang: Hepatitic C merupakan penyakit yang disebabkan oleh Virus Hepatitis C (HCV) dan dapat mengakibatkan peradangan hati, biasanya bersifat asimtomatik, bahkan kronik yang ditandai dengan sirosis, kanker hati, kelainan fungsi hati yang dapat menyebabkan kematian. Pengobatan standar dengan PEGinterferon-a dan ribavirin memiliki efek samping yang berat, sehingga diperlukan pengobatan alternatif sebagai anti-HCV. Dimocarpus longan merupakan tanaman yang memiliki khasiat sebagai antijamur, antivirus, antiinflamasi, antioksidan, antibakteri, dan antikanker. Tujuan penelitian ini untuk mengetahui efek dan mekanisme kerja ekstrak metanol daun D. longan terhadap virus Hepatitis C.Metode: Cell line derivate Human hepatocarcinoma (Huh 7it-1) diinfeksikan dengan HCV strain JFH1 dari genotipe 2a yang diinduksikan ekstrak metanol daun D. longan selama dua hari, kemudian diukur virus yang terbentuk ekstraseluler dan intraseluler. Pemeriksaan virus ekstraseluler dengan cara focus forming assay sedangkan intraseluler dengan qRT-PCR, western blot dan relative fluorescence assay. Pengujian sitotoksisitas terhadap Huh 7it-1 dengan metode MTT assay. Ekstrak metanol daun D. longan diuji adanya kandungan saponin, flavonoid, triterpenoid dan steroid, tanin, dan glikosida.Hasil: Konsentrasi hambatan 50% (IC50) ekstrak terhadap HCVsebesar 13,2 ± 0,52 μg/ml dan toksik 50% (CC50) terhadap sel Huh 7it-1 sebesar 681,9 ± 13,2 μg/ml dengan nilai indek selektivitas (SI) sebesar 51,2. Efek virusidal ekstrak metanol daun D. longan secara langsung terhadap HCV berupa pengurangan titer virus sebesar 99%. Analisis RNA dan protein NS3 HCV intraseluler memperlihatkan adanya hambatan sebesar 20%. Kandungan fitokimia yang terdapat pada ekstrak metanol daun D. longan di antaranya saponin, flavonoid, triterpenoid dan steroid, alkaloid, tanin, dan glikosida.Kesimpulan: Mekanisme anti-HCV dari ekstrak metanol daun D. longan diduga melalui adanya hambatan pada entry dan post-entry yang bekerja dengan menghambat pada penempelan virus, membunuh virus dengan interaksi langsung, menghambat ekspresi NS3, dan menghambat replikasi. Kandungan fitokimia yang terkandung seperti saponin, flavonoid, triterpenoid dan steroid, tanin, dan glikosida.

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Background: Hepatitic C is a disease caused by the hepatitis C virus (HCV). HCV infection can lead to inflammation of liver tend to be asymptomatic, and chronic characterized by cirrhosis, liver cancer, abnormal liver function can cause mortality. Standard HCV treatment with PEG-interferon-a and ribavirin have severe side effects, necessitating alternative treatments as anti-HCV. Dimocarpus longan is a plant that previously reported has antifungal, antiviral, anti-inflammatory, antioxidant, antibacterial, and anticancer activity. The purpose of this study is determine the effects and mechanism of action of the methanol extract of leaves of D. longan against hepatitis C virus.Methods: A derivate of Human hepatocarcinoma Cell line (Huh 7it-1) was infected with HCV of genotype 2a JFH1 strain which is inducted with methanol extracts of D. longan for two days and then number of virus produced outside of the cell (extracellular) and inside of the cell (intracellular) were measured by focus forming assay, while intracellular virus was measured by qRT-PCR, western blot and relative fluorescence assay. Cytotoxicity against Huh 7it-1 was tested by MTT assay. Examination of phytochemical content D. longan showed the presence of saponins, flavonoids, triterpenoids and steroids, alkaloids, tannins, and glycosides.Result: D. longan concentration of inhibition 50% (IC50) and Toxic effects of concentration of cytotoxicity 50% (CC50) againts cells Huh 7it-1were obtained 13,2 ± 0.52 ug/ml and 681.9 ± 13.2 ug/ml, respectively and with selectivity index (SI) 51.2. Result of direct virucidal effect was shown inhibition of titer virus 99%. RNA and NS3 protein analysis of HCV were shown inhibition 20%. Phytochemical contains of methanol extracts of Dimocarpus longan Lour. Leaves are saponins, flavonoids, triterpenoids and steroids, tannins, and glycosides.Conclusion: Anti-HCV mechanisms of methanol extracts of Dimocarpus longan Lour. Leaves are inhibition at entry and post-entry with action at attachment, direct killing, inhibition of expression NS3, and replication. Phytochemical content in the methanol extract of leaves of D. longan were saponins, flavonoids, triterpenoids and steroids, tannins, and glycosides."

"ABSTRAKHepatitis B adalah suatu penyakit yang disebabkan oleh virus hepatitis B. Salah satu faktor yang berpengaruh dalam pembentukan virus hepatitis B adalah core protein Cp . Sehingga Cp dapat digunakan sebagai salah satu target pengobatan hepatitis B. Pada penelitian ini dilakukan penapisan virtual senyawa dari basis data tanaman herbal Indonesia sebagai core protein allosteric modulator CpAM menggunakan peranti lunak AutoDock dan AutoDock Vina. Metode divalidasi dengan menggunakan parameter Enrichment Factor EF , Receiver Operating Characteristics ROC , dan Area Under Curve AUC . Pada penapisan menggunakan AutoDock digunakan grid box ukuran 55x55x55 dengan nilai EF10 0.7652 dan AUC 0.6709 sementara grid box ukuran 20.625x20.625x20.625 untuk penapisan menggunakan AutoDock Vina dengan nilai EF5 0.5075 dan AUC 0.7832. Sepuluh senyawa terbaik hasil penapisan virtual menggunakan AutoDock memiliki rentang DG: -11.74 -10.31 kkal/mol adalah yuehchukene, lansionic acid, stigmast-4-en-3-one, myrtillin, sanggenol O, lanosterol, erycristagallin, alpha-spinasterol, cyanidin 3-arabinoside, dan cathasterone. Sepuluh senyawa terbaik hasil penapisan virtual menggunakan AutoDock Vina memiliki rentang DG: -12.1 -10.7 kkal/mol adalah sanggenol O, cucumerin A, yuehchukene, palmarumycin CP1, dehydrocycloguanandin, myrtilin, liriodenine, myricetin 3-alpha-L-Arabinopyranoside, myricetin 3-galactoside, dan cassameridine.

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ABSTRACTHepatitis B is a disease caused by hepatitis B virus. One of the main factor in virus assembly is core protein Cp . Therefore Cp is suitable to use as one of therapeutic target for hepatitis B. In this study virtual screening of Indonesia herbal database as CpAM of hepatitis B virus was performed using AutoDock and AutoDock Vina software. The methode was validated by Enrichment Factor EF , Receiver Operating Characteristics ROC , and Area Under Curve AUC parameters. The grid box size used in virtual screening with AutoDock is 55x55x55 with EF10 0.7652 and AUC 0.6709 meanwhile grid box size that will be use in virtual screening using AutoDock Vina is 20.625x20.625x20.625 with EF5 0.5075 and AUC 0.7832. The best top ten compounds from virtual screening with AutoDock has DG levels 11.74 10.31 kkal mol theare yuehchukene, lansionic acid, stigmast 4 en 3 one, myrtillin, sanggenol O, lanosterol, erycristagallin, alpha spinasterol, cyanidin 3 arabinoside, dan cathasterone. The best top ten compounds from virtual screening with AutoDock Vina has DG levels 12.1 10.7 kkal mol adalah sanggenol O, cucumerin A, yuehchukene, palmarumycin CP1, dehydrocycloguanandin, myrtilin, liriodenine, myricetin 3 alpha L Arabinopyranoside, myricetin 3 galactoside, dan cassameridine"

"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."

"Latar belakang: Penelitian ini bertujuan untuk merancang dan menganalisis pelacak oligonukleotida apakah dapat diterapkan dalam hibridisasi dot blot menggunakan radioisotop 32P untuk mendeteksi virus hepatitis C.Metode: Sampel yang digunakan adalah 46 plasma darah. Plasma diekstraksi untuk mendapatkan RNA genom virus sebagai cetakan reaksi RT-PCR dan amplikon digunakan untuk nested PCR. Genom HCV berjumlah 24 diunduh dari GeneBank dan penderetan sekuen DNA dilakukan dengan Software Bio Edit versi 7.0.9.0. Pelacak oligonuklueotida dirancang berdasarkan daerah lestari genom HCV yang terletak pada sekuen internal di antara 2 primer yang digunakan pada nested PCR. Homologi oligonukleotida HCV dianalisis menggunakan teknik Blast di GeneBank. Radioisotop 32P digunakan untuk melabel oligonukleotida. Oligonukleotida berlabel diaplikasikan untuk produk nested PCR menggunakan metode hibridisasi dot blot. Konfirmasi hasil amplifikasi dan hibridisasi dot blot dilakukan menggunakan metode sekuensing DNA.Hasil: Hasil analisis Blast menunjukkan homologi yang tinggi untuk HCV (100%). Hasil nested PCR menunjukkan tiga pola fragmen DNA. Tiga pola tersebut masing-masing adalah genotip HCV 1, 2, dan 3. Primer yang digunakan dalam nested PCR tidak spesifik dinyatakan dengan adanya tiga fragmen DNA sehingga sulit diinterpretasikan. Hasil hibridisasi dot blot menggunakan oligonukleotida yang didesain dalam penelitian ini menunjukkan intensitas dot yang tebal. Semua pola fragmen hasil nested PCR menunjukkan hasil positif dot blot. Hasil hibridisasi dot blot sesuai dengan hasil sekuensing DNA.Kesimpulan: Pelacak oligonukleotida menunjukkan kriteria yang sangat memuaskan secara bioinformatika. Hasil hibridisasi dot blot menggunakan 32P menunjukkan intensitas dot yang tebal dan lebih mudah diinterpretasi dibandingkan dengan hasil nested PCR.

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AbstractBackground: This study aimed to design and analyze the applicability of an oligonucleotide probe in radioisotope 32P-based dot blot hybridization for detection of hepatitis C virus.Methods: Forty-six of plasma samples were used. The plasma was extracted to obtain viral RNA genome as template for RT-PCR and the amplicon was used for nested PCR. Twenty-four HCV genomes were retrieved from GeneBank DNA sequence and alignment was performed by Bio Edit Software version 7.0.9.0. An oligonucleotide probe was designed based on a highly conserved region that is located on internal sequence between two primers used for nested PCR. Blast analysis on GeneBank was performed to obtain homology of the oligonucleotide for HCV. The oligonucleotide was then labeled with 32P and dot blot hybridization was applied for nested PCR products. DNA Sequencing was performed to confirm the amplicon and dot blot hybridization results.Results: Blast analysis showed high homology (100%) for HCV. Nested PCR resulted in three patterns of DNA fragments representing HCV genotypes 1, 2, and 3, respectively. The primers used in nested PCR were not specific and resulted in DNA fragments difficult to be interpreted. Dot blot hybridization using the designed oligonucleotide showed high intensity dots. All nested PCR fragments showed the dot blot positive. The dot blot results were in accordance with DNA sequencing that confirmed three patterns of DNA fragments as different HCV genotypes.Conclusion: The oligonucleotide showed excellent bioinformatically criteria. 32P-based dot blot hybridization yielded high intensity dots and was easier to be interpreted than nested PCR assay."

"[ABSTRAKStudi cross sectional pada pasien hepatitis B di Indonesia menunjukkan korelasi mutasi kodon start pre-S2 dengan keparahan penyakit hati. Peran protein-protein HBs pada aktivasi NF-ĸB sebagai salah satu faktor dalam induksi keparahan penyakit hati. Studi ini dilakukan untuk melihat efek varian mutan HBs virus hepatitis B subgenotipe B3 sebagai strain endemik di Indonesia pada keparahan penyakit hati dilihat dari ekspresi dan aktivasi NF-ĸB. Gen HBs dari tiga pasien yang membawa tiga varian HBs berbeda diamplifikasi dan diklon dengan plasmid pcDNA3.1, ditransfeksikan dengan metode lipofektamin ke dalam sel Huh7. Nilai ekspresi mRNA dianalisis dengan real-time PCR terhadap mRNA HBs, IĸB-α, dan NF-ĸB (p50). Ekspresi IĸB-α yang diregulasi oleh NF-ĸB digunakan sebagai parameter untuk aktivasi NF-ĸB. Diperoleh plasmid ekspresi HBs dengan mutasi kodon start pre-S2, delesi pre-S2 dan wild type VHB subgenotipe B3. Plasmid rekombinan pcDNA HBs dapat mengekspresikan mRNA HBs dan menurun pada 48 hingga 72 jam. Kecuali pada mutan delesi pre-S2 yang stabil hingga 72 jam. Ekspresi protein HBs berdasar ELISA menunjukkan nilai relatif konstan pada HBs wild type, sedangkan pada HBs mutan kodon start dan delesi meningkat pada 72 jam. Aktivasi NF-ĸB relatif lebih tinggi oleh tipe wild type dibanding mutan kodon start pre-S2 dan delesi pre-S2, sehingga variasi mutasi tidak memberikan pengaruh pada aktivasi NF-ĸB, meski varian mutan delesi pre-S2 menunjukkan peningkatan aktivasi NF-ĸB setelah waktu kultur yang lebih lama dibanding HBs wild type dan mutan kodon start pre-S2. Ekspresi NF-ĸB (p50) dipengaruhi oleh variasi mutasi, ekspresi p50 lebih tinggi pada mutan kodon start pre-S2 dibanding varian HBs lainnya. Keparahan penyakit hati oleh mutasi kodon start pre-S2 dapat terkait dengan peningkatan ekspresi p50.

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ABSTRACTCross sectional study on hepatitis B patients in Indonesia showed association of pre-S2 start codon mutation with severity liver disease. Role of HBs proteins on the activation of NF-ĸB as one of the factor in liver disease progression. This study was to see the effects of different HBs mutant variants of Hepatitis B Virus (HBV) subgenotype B3 as the endemic strain in Indonesia on the expression and activation of NF-ĸB. HBs genes of three hepatitis B patients were amplified and cloned to pcDNA3.1, and were transfected using lipofectamine into Huh7 cell line. Expressions on mRNA level for HBs, IĸB-α and NF-ĸB (p50) were evaluated using real-time PCR. IĸB-α expression which is regulated by NF-ĸB was used as parameter to measure NF-ĸB activation. Recombinant plasmid for HBs expression with pre-S2 start codon mutation, pre-S2 deletion and wild type of HBV subgenotipe B3 were obtained. All three clones showed high level of mRNA expression which decreased after 48 to 72 hours, except for pre-S2 deletion which was relatively stabil up to72 hours. HBs protein expression detected using ELISA was constant for HBs wild type whilst increased at 72 hours for pre-S2 start codon mutation and pre-S2 deletion. NF-ĸB activation was higher for HBs wild type compared to the two mutant variants, suggesting no effect of mutation to increment of NF-ĸB activation, however pre-S2 deletion mutant showed higher NF-ĸB activation after longer period of incubation. NF-ĸB (p50) expression was higher for pre-S2 start codon mutation, suggesting liver disease progression by pre-S2 start codon mutation might associated to increased expression of p50.;Cross sectional study on hepatitis B patients in Indonesia showed association of pre-S2 start codon mutation with severity liver disease. Role of HBs proteins on the activation of NF-ĸB as one of the factor in liver disease progression. This study was to see the effects of different HBs mutant variants of Hepatitis B Virus (HBV) subgenotype B3 as the endemic strain in Indonesia on the expression and activation of NF-ĸB. HBs genes of three hepatitis B patients were amplified and cloned to pcDNA3.1, and were transfected using lipofectamine into Huh7 cell line. Expressions on mRNA level for HBs, IĸB-α and NF-ĸB (p50) were evaluated using real-time PCR. IĸB-α expression which is regulated by NF-ĸB was used as parameter to measure NF-ĸB activation. Recombinant plasmid for HBs expression with pre-S2 start codon mutation, pre-S2 deletion and wild type of HBV subgenotipe B3 were obtained. All three clones showed high level of mRNA expression which decreased after 48 to 72 hours, except for pre-S2 deletion which was relatively stabil up to72 hours. HBs protein expression detected using ELISA was constant for HBs wild type whilst increased at 72 hours for pre-S2 start codon mutation and pre-S2 deletion. NF-ĸB activation was higher for HBs wild type compared to the two mutant variants, suggesting no effect of mutation to increment of NF-ĸB activation, however pre-S2 deletion mutant showed higher NF-ĸB activation after longer period of incubation. NF-ĸB (p50) expression was higher for pre-S2 start codon mutation, suggesting liver disease progression by pre-S2 start codon mutation might associated to increased expression of p50.;Cross sectional study on hepatitis B patients in Indonesia showed association of pre-S2 start codon mutation with severity liver disease. Role of HBs proteins on the activation of NF-ĸB as one of the factor in liver disease progression. This study was to see the effects of different HBs mutant variants of Hepatitis B Virus (HBV) subgenotype B3 as the endemic strain in Indonesia on the expression and activation of NF-ĸB. HBs genes of three hepatitis B patients were amplified and cloned to pcDNA3.1, and were transfected using lipofectamine into Huh7 cell line. Expressions on mRNA level for HBs, IĸB-α and NF-ĸB (p50) were evaluated using real-time PCR. IĸB-α expression which is regulated by NF-ĸB was used as parameter to measure NF-ĸB activation. Recombinant plasmid for HBs expression with pre-S2 start codon mutation, pre-S2 deletion and wild type of HBV subgenotipe B3 were obtained. All three clones showed high level of mRNA expression which decreased after 48 to 72 hours, except for pre-S2 deletion which was relatively stabil up to72 hours. HBs protein expression detected using ELISA was constant for HBs wild type whilst increased at 72 hours for pre-S2 start codon mutation and pre-S2 deletion. NF-ĸB activation was higher for HBs wild type compared to the two mutant variants, suggesting no effect of mutation to increment of NF-ĸB activation, however pre-S2 deletion mutant showed higher NF-ĸB activation after longer period of incubation. NF-ĸB (p50) expression was higher for pre-S2 start codon mutation, suggesting liver disease progression by pre-S2 start codon mutation might associated to increased expression of p50., Cross sectional study on hepatitis B patients in Indonesia showed association of pre-S2 start codon mutation with severity liver disease. Role of HBs proteins on the activation of NF-ĸB as one of the factor in liver disease progression. This study was to see the effects of different HBs mutant variants of Hepatitis B Virus (HBV) subgenotype B3 as the endemic strain in Indonesia on the expression and activation of NF-ĸB. HBs genes of three hepatitis B patients were amplified and cloned to pcDNA3.1, and were transfected using lipofectamine into Huh7 cell line. Expressions on mRNA level for HBs, IĸB-α and NF-ĸB (p50) were evaluated using real-time PCR. IĸB-α expression which is regulated by NF-ĸB was used as parameter to measure NF-ĸB activation. Recombinant plasmid for HBs expression with pre-S2 start codon mutation, pre-S2 deletion and wild type of HBV subgenotipe B3 were obtained. All three clones showed high level of mRNA expression which decreased after 48 to 72 hours, except for pre-S2 deletion which was relatively stabil up to72 hours. HBs protein expression detected using ELISA was constant for HBs wild type whilst increased at 72 hours for pre-S2 start codon mutation and pre-S2 deletion. NF-ĸB activation was higher for HBs wild type compared to the two mutant variants, suggesting no effect of mutation to increment of NF-ĸB activation, however pre-S2 deletion mutant showed higher NF-ĸB activation after longer period of incubation. NF-ĸB (p50) expression was higher for pre-S2 start codon mutation, suggesting liver disease progression by pre-S2 start codon mutation might associated to increased expression of p50.]"