Locked nucleic acid real-time polymerase chain reaction method identifying two polymorphisms of hepatitis B virus genotype C2 infections,rt269L and rt269I  

作　　者：Kijeong Kim Yu-Min Choi Dong Hyun Kim Junghwa Jang Won Hyeok Choe Bum-Joon Kim 

机构地区：[1]Department of Microbiology,College of Medicine,Chung-Ang University,Seoul 06974,South Korea [2]Department of Microbiology and Immunology,College of Medicine,Seoul National University,Seoul 03080,South Korea [3]Department of Internal Medicine,Konkuk University School of Medicine,Seoul 05030,South Korea [4]Department of Biomedical Sciences,College of Medicine,Seoul National University,Seoul 03080,South Korea [5]Liver Research Institute,College of Medicine,Seoul National University,Seoul 03080,South Korea [6]Seoul National University Medical Research Center,Seoul 03080,South Korea

出　　处：《World Journal of Gastroenterology》2023年第11期1721-1734,共14页世界胃肠病学杂志（英文版）

基　　金：Supported by the National Research Foundation of Korea,No.2022R1A2B5B01001421;the Korea Health Technology R&D Project through the Korea Health Industry Development Institute,the Ministry of Health&Welfare,Republic of Korea,No.HI22C0476.

摘　　要：BACKGROUND The presence of two distinct hepatitis B virus(HBV)Pol RT polymorphisms,rt269L and rt269I,could contribute to the unique clinical or virological phenotype of HBV genotype C2.Therefore,a simple and sensitive method capable of identifying both types in chronic hepatitis B(CHB)patients infected with genotype C2 should be developed.AIM To develop a novel simple and sensitive locked nucleic acid(LNA)-real timepolymerase chain reaction(RT-PCR)method capable of identifying two rt269 types in CHB genotype C2 patients.METHODS We designed proper primer and probe sets for LNA-RT-PCR for the separation of rt269 types.Using synthesized DNAs of the wild type and variant forms,melting temperature analysis,detection sensitivity,and endpoint genotyping for LNA-RT-PCR were performed.The developed LNA-RT-PCR method was applied to a total of 94 CHB patients of genotype C2 for the identification of two rt269 polymorphisms,and these results were compared with those obtained by a direct sequencing protocol.RESULTS The LNA-RT-PCR method could identify two rt269L and rt269I polymorphisms of three genotypes,two rt269L types[‘L1’(WT)and‘L2’]and one rt269I type(‘I’)in single(63 samples,72.4%)or mixed forms(24 samples,27.6%)in 87(92.6%sensitivity)of 94 samples from Korean CHB patients.When the results were compared with those obtained by the direct sequencing protocol,the LNA-RT-PCR method showed the same results in all but one of 87 positive detected samples(98.9%specificity).CONCLUSION The newly developed LNA-RT-PCR method could identify two rt269 polymorphisms,rt269L and rt269I,in CHB patients with genotype C2 infections.This method could be effectively used for the understanding of disease progression in genotype C2 endemic areas.

关 键 词：Hepatitis B virus Genotype C2 POLYMERASE rt269 Locked nucleic acid-real time-polymerase chain reaction Chronic hepatitis B 

分 类 号：R512.62[医药卫生—内科学]