期刊论文

Development and evaluation of real-time fluorescence recombinase aided amplification assay without extracting nucleic acid for detection of adenovirus type 3

中文

中华实验和临床病毒学杂志

1003-9279

2019

33

6

653-657

10.3760/cma.j.issn.1003-9279.2019.06.020

2018ZX10711001:十三五重大项目 2017ZX10104001:十三五重大项目 2018ZX10713-002:十三五重大项目 2017SK2101:湖南省重点研发计划

本校为第一机构

目的 建立3型腺病毒(human adenovirus 3,HAdV-3)免提取核酸的重组酶介导的等温扩增(recmbinase acid amplification,RAA)实时荧光检测方法。方法 根据HAdV-3基因保守序列分别设计1对引物和1条探针,通过对免提核酸的条件摸索和优化,建立1种免提取核酸,重组酶介导的等温扩增实时荧光检测方法;通过对培养的毒株进行系列稀释,免提取核酸分析该方法的灵敏度;通过检测其他呼吸道病毒的原始样本,评价该方法的特异性;并检测HAdV-3临床样本,与传统的提核酸实时荧光定量PCR方法进行比较。结果 本研究建立的免提取核酸实时荧光RAA方法对10倍系列稀释的HAdV-3毒株进行检测,和实时荧光定量PCR方法相比,灵敏度...

Objective To establish a real-time fluorescence recombinase acid amplification (RAA)method for the detection of adenovirus type 3 (HAdV-3)without extraction nucleic acid. Methods According to the conserved sequence of adenovirus type 3 gene, a pair of primers and a probe were designed, and a real-time fluorescence RAA without extracting nucleic acid was established and optimizing the condition of DNA-free extraction. The sensitivity of the method was analyzed by a series of dilution and the specificity of the method was evaluated by detecting the original samples of other respiratory viruses. ...