Accurate genotyping of hepatitis C virus through nucleotide sequencing and identification of new HCV subtypes in China population
Accurate genotyping of hepatitis C virus through nucleotide sequencing and identification of new HCV subtypes in China population
复制标题
通过核苷酸测序对丙型肝炎病毒进行准确的基因分型并鉴定中国人群中的新的丙型肝炎病毒亚型
DOI：
10.1016/j.cmi.2015.05.034
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发表时间：
2015-09-01
影响因子：
14.2
通讯作者：
Li, Y.
中科院分区：
文献类型：
--
作者：
Tong, Y. -Q.;Liu, B.;Li, Y.
NS 5 B基因组信息区的核苷酸序列测定仍然是丙型肝炎病毒（HCV）基因分型的金标准。在这里，我们开发了一种新的方法来测序新的NS 5 B区域，以提高HCV基因分型和亚型的准确性和灵敏度。通过对HCV数据库中1127条HCV基因组全长NS 5 B区序列进行扫描，筛选出8条新引物。对每对引物扩增HCV亚型的能力进行评分，新引物能够比以前使用的引物更好地扩增NS 5 B区域，因此更准确地分型HCV毒株。用新引物对扩增的DNA进行测序，可以特异、准确地检测出HCV的5种标准亚型（1a、2a、3b、6a和1b）。我们进一步检查了患者样本，发现新引物能够以高灵敏度鉴定临床样本中的HCV亚型。该方法能检测567份临床标本中的所有亚型HCV。其中发现了3种新的HCV亚型（1b-2a、1b-2k和6d-6 k），在国内未见报道。总之，测序的NS 5 B区域扩增的新的NS 5 B引物是一个更可靠的方法，HCV基因分型和更敏感的诊断工具，比测序使用先前描述的引物，并可以识别新的HCV亚型。本研究对HCV的临床诊断、指导临床治疗、患者管理及流行病学研究具有重要意义。临床微生物学和感染（C）2015年欧洲临床微生物学和传染病学会。由爱思唯尔有限公司出版。保留所有权利。
Nucleotide sequencing of the phylogenetically informative region ofNS5B remains the gold standard for hepatitis C virus (HCV) genotyping. Here we developed a new methodology for sequencing newNS5B regions to increase the accuracy and sensitivity ofHCV genotyping and subtyping. The eight newprimers were identified by scanning the full-length NS5B regions from 1127 HCV genomic sequences found in HCV databases. The ability of each pair of primers to amplify HCV subtypes was scored, and the new primers were able to amplify the NS5B region better than the previously used primers, therefore more accurately subtyping HCV strains. Sequencing the DNA amplified by the new primer pairs can specifically and correctly detect the five standard HCV subtypes (1a, 2a, 3b, 6a and 1b). We further examined patient samples and found that the new primers were able to identify HCV subtypes in clinical samples with high sensitivity. This method was able to detect all subtypes of HCV in 567 clinical samples. Importantly, three novel HCV subtypes (1b-2a, 1b-2k and 6d-6k) were identified in the samples, which have not been previous reported in China. In conclusion, sequencing the NS5B region amplified by the new NS5B primers is a more reliable method of HCV genotyping and a more sensitive diagnostic tool than sequencing using the previously described primers, and could identify new HCV subtypes. Our research is useful for clinical diagnosis, guidance of clinical treatment, management of clinical patients, and studies on the epidemiology ofHCV. Clinical Microbiology and Infection (C) 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.