A novel hepatitis B virus genotyping system by using restriction fragment length polymorphism patter

来源 :World Journal of Gastroenterology | 被引量 : 0次 | 上传用户:xpz_peng
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AIM:Traditional hepatitis B virus(HBV)genotyping methodsusing restriction fragment length polymorphism(RFLP)canreliably identify genotypes A to F.As HBV genotypes G andH have been recently identified,this study was to establishan accurate and simple genotyping method for all eightHBV genotypes(A to H).METHODS:Two hundred and forty HBV small S sequencesobtained from GeneBank were analysed for restrictionenzyme sites that would be genotype-specific.Restrictionpatterns following digestion with restriction enzymes BsrIⅠStyⅠ,DpnⅠ,Hpa/Ⅱ,and EaeⅠ,were determined to identify alleight HBV genotypes.Mixed genotype infections wereconfirmed by cloning and further RFLP analysis.RESULTS:The new genotyping method could identify HBVgenotypes A to H.Genotypes B and C could be determined bya single step digestion with BsrⅠ and StyⅠ in parallel.This wasparticularly useful in the Far East where genotypes B and C arepredominant.Serum samples from 187 Chinese HBV carrierswere analysed with this genotyping system,and the genotypedistnbution was 1.1%(2),51.9%(97),40.6%(76)and 4.8%(9)for genotypes A,B,C,and D,respectively.Mixed genotypeswere found in only 3 patients(1.6%).Sequence data analysisconfirmed the validity of this new method.CONCLUSION:This HBV genotyping system can identifyall eight HBV genotypes.It is accurate and simple,and canbe widely used for studies on HBV genotyping.Zeng GB,Wen SJ,Wang ZH,Yan L,Sun J,Hou JL.A novelhepatitis B virus genotyping system by using restrictionfragment length polymorphism patterns of S gene amplicons.World J Gastroenterol 2004,10(21):3132-3136http://www.wjgnet.com/1007-9327/10/3132.asp AIM: Traditional hepatitis B virus (HBV) genotyping methods using restriction fragment length polymorphism (RFLP) canreliably identify genotypes A to F.As HBV genotypes G andH have been recently identified, this study was to establishan accurate and simple genotyping method for all eight HBV genotypes ( A to H) .METHODS: Two hundred and forty HBV small S sequence derived from GeneBank were analyzed for restriction enzyme sites that would be genotype-specific. Restriction patterns following digestion with restriction enzymes BsrI I StyI, DpnI, Hpa / II, and EaeI, were determined to identify alleight HBV genotypes. Mixed genotype infections were confirmed by cloning and further RFLP analysis. RESULTS: The new genotyping method could identify HBV genotypes A to H. Genotypes B and C could be determined by a single step digestion with BsrI and StyI in parallel. This wasparticularly useful in the Far East where genotypes B and C are predominant. Serum samples from 187 Chinese HBV carrierswere analysed with thi s genotyping system and genotypedistnbution was 1.1% (2), 51.9% (97), 40.6% (76) and 4.8% (9) for genotypes A, B, C and D, respectively. patients (1.6%). Sequence data analysis confirmed the validity of this new method. CONCLUSION: This HBV genotyping system can identify all eight HBV genotypes. Is accurate and simple, and can widely used for studies on HBV genotyping. Zeng GB, Wen SJ, Wang ZH, Yan L, Sun J, Hou JL. A novel hepatitis B virus genotyping system by using restriction fragment length polymorphism patterns of S gene amplicons. World J Gastroenterol 2004, 10 (21): 3132-3136 http: //www.wjgnet.com /1007-9327/10/3132.asp
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