PCR-Based In Vitro Synthesis of Hepatitis C Virus NS3 Protease for Rapid Phenotypic Resistance Testing of Protease Inhibitors

doi:10.1128/JCM.03257-13

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	PCR-Based In Vitro Synthesis of Hepatitis C Virus NS3 Protease for Rapid Phenotypic Resistance Testing of Protease Inhibitors
	Qiao, Jinjuan 1,2,3; Yu, Junping 1,2; Yang, Hang 1,2; Wei, Hongping 1,2
刊名	JOURNAL OF CLINICAL MICROBIOLOGY
	2014-04-01
卷号	52 期号:4 页码:1139-1145
ISSN号	0095-1137
DOI	10.1128/JCM.03257-13
英文摘要	Protease inhibitors (PIs) targeting the hepatitis C virus (HCV) NS3 protease, such as telaprevir, have significantly improved the sustained virologic response (SVR) rates of HCV genotype 1 antiviral therapy. Given the expanding antiviral therapy regimen, fast HCV PI resistance assays are urgently needed. In this view, we have developed a novel phenotypic resistance test for HCV PIs based on in vitro synthesis of patient-derived HCV NS3 protease and subsequent enzymatic testing in a fluorescent readout. The enzymatically active HCV NS3 proteases were synthesized from PCR-derived templates by an Escherichia coli S30 extract system. Tests of the protease genes with known mutations for telaprevir resistance showed that the phenotypic resistance test was fast, with a total turnaround time of <10 h, and was fully in agreement with the previous resistance results. The initial tests with 38 treatment-naive serum samples showed that the method was significantly less laborious and faster than currently available phenotypic resistance assays of HCV NS3 PIs.
资助项目	National Natural Science Foundation of China[21105114] ; National Natural Science Foundation of China[21075131] ; Key Laboratory on Emerging Infectious Diseases and Biosafety in Wuhan, China
WOS研究方向	Microbiology
语种	英语
出版者	AMER SOC MICROBIOLOGY
WOS记录号	WOS:000332832300019
内容类型	期刊论文
源URL	[http://202.127.146.157/handle/2RYDP1HH/3931]
专题	中国科学院武汉植物园
通讯作者	Wei, Hongping
作者单位	1.Chinese Acad Sci, Ctr Emerging Infect Dis, Wuhan, Peoples R China 2.Chinese Acad Sci, State Key Lab Virol, Wuhan Inst Virol, Wuhan, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Qiao, Jinjuan,Yu, Junping,Yang, Hang,et al. PCR-Based In Vitro Synthesis of Hepatitis C Virus NS3 Protease for Rapid Phenotypic Resistance Testing of Protease Inhibitors[J]. JOURNAL OF CLINICAL MICROBIOLOGY,2014,52(4):1139-1145.
APA	Qiao, Jinjuan,Yu, Junping,Yang, Hang,&Wei, Hongping.(2014).PCR-Based In Vitro Synthesis of Hepatitis C Virus NS3 Protease for Rapid Phenotypic Resistance Testing of Protease Inhibitors.JOURNAL OF CLINICAL MICROBIOLOGY,52(4),1139-1145.
MLA	Qiao, Jinjuan,et al."PCR-Based In Vitro Synthesis of Hepatitis C Virus NS3 Protease for Rapid Phenotypic Resistance Testing of Protease Inhibitors".JOURNAL OF CLINICAL MICROBIOLOGY 52.4(2014):1139-1145.