中国兽医科学2018，Vol.48Issue(5)：552-558,7.DOI:10.16656/j.issn.1673-4696.2018.0103
蓝舌病病毒10型和20型及23型多重RT-PCR检测方法的建立
Establishment of multiplex RT-PCR for the detection of bluetongue virus types 10, 20 and 23
摘要
Abstract
To develop an assay for the simultaneous identification of bluetongue virus (BTV) 10/ 20/23, a multiplex RT-PCR was established with 3 pairs of specific primers targeting the sequence of BTV VP2 gene. Through optimizing the reaction conditions,the multiplex RT-PCR was able to specifically detect BTV type 10,20 and 23 simultaneously with the detection limit of 1.696×103 copies/μL for type 10,1.375×103copies /μL for type 20 and 1.317×103 copies/μL for type 23, respectively, but no genomic RNA or DNA were amplified from the other BTV genotype,peste des ptits ruminants virus (PPRV)and foot-and-mouth disease virus(FMDV). Using the assay to detect 67 clinical samples,the results showed that the coincidence between the n-PCR of OIE and the multiplex RT-PCR was 100％. In conclusion,the established multiplex RT-PCR in this study was specific and sensitive for the clinical samples,and could identify the serotype quickly and exceptionally.关键词
蓝舌病病毒/多重RT-PCR/血清型鉴别Key words
BTV/multiplex RT-PCR/serotype identification分类
农业科技引用本文复制引用
黄秋华,王国民,胡世君,聂福平,周庆,杨俊,王昱,艾军,唐昌杰,陈冉越,李应国..蓝舌病病毒10型和20型及23型多重RT-PCR检测方法的建立[J].中国兽医科学,2018,48(5):552-558,7.基金项目
重庆市重大应用开发项目(cstc,2015yykfC80001) （cstc,2015yykfC80001）
质检公益性行业科研专项(201310093) （201310093）
