鸡腺病毒血清4型变异株荧光定量PCR检测方法的建立  

Establishment of real-time quantitative PCR method for detection of fowl adenovirus serotype 4 mutant strain

作　　者：陈媛[1] 屈贵蜀 彭尧舜 黄瑞玲 许丽惠[1] 王全溪[1] CHEN Yuan;QU Gui-shu;PENG Yao-shun;HUANG Rui-ling;XU Li-hui;WANG Quan-xi(College of Animal Sciencey Fujian Agriculture and Forestry Universityf Fuzhou 350002,China)

机构地区：[1]福建农林大学动物科学学院

出　　处：《中国兽医科学》2019年第8期1013-1018,共6页Chinese Veterinary Science

基　　金：福建省自然科学基金项目(2019J01658);福建农林大学科技发展基金项目(CXZX201789)

摘　　要：为建立一种快速、灵敏的Ⅰ群鸡腺病毒血清4型(FAdV4)变异株检测方法,针对FAdV4变异株NP株Hexon基因的保守序列设计1对特异引物,通过荧光定量PCR(QPCR)反应体系的优化,建立了FAdV4的QPCR检测方法。结果显示:该方法特异性强,除FAdV4变异株外均无其他非特异扩增;此外,该方法敏感度高,其检测下限为53.9 copies/μL;组内、组间变异系数均小于5%,重复性好。临床样品检测结果显示,建立的QPCR方法的FAdV4变异株检出率为56.5%,远高于常规PCR的检出率(21.7%)。可见,建立的QPCR方法可用于临床快速检测FAdV4变异株。In order to establish a rapid and sensitive method for detection on fowl adenovirus serotype 4(FAdV4) mutant strain, a pair of specific primers was designed based on the conserved sequence of Hexon gene of NP strain of FAdV4.The QPCR reaction condition was optimized.The results showed that the QPCR method for FAdV4 was highly specific.In addition,the method had high sensitivity because the detection limit was to 53.9 copies/μL.The variation coefficients of intra-and inter-assay were less than 5%.The results of samples detection showed that the positive rate of FAdV4 mutant strain was to 56.5% detected by the QPCR method,which was higher than that of conventional PCR(21.7%).Therefore,the QPCR method can be used for rapid detection of FAd V4 mutant strain.

关 键 词：鸡腺病毒血清4型 变异株 荧光定量PCR 诊断方法 Hexon基因 

分 类 号：S852.659.1[农业科学—基础兽医学]