Development and evaluation of real-time fluorescence recombinase aided amplification assay without extracting nucleic acid for detection of adenovirus type 3
10.3760/cma.j.issn.1003-9279.2019.06.020
- VernacularTitle: 3型腺病毒免提取核酸重组酶介导的等温扩增实时荧光检测方法的建立及应用
- Author:
Ruihua WANG
1
,
2
;
Yi ZHANG
3
;
Xingyu XIANG
1
;
Zhifei ZHAN
4
;
Xinna LI
3
;
Xinxin SHEN
3
;
Zhen ZHU
3
;
Ruiqing ZHANG
3
;
Xueding BAI
3
;
Qingxia DUAN
3
;
Guohao FAN
3
;
Hong ZHANG
1
;
Xuejun MA
3
Author Information
1. Hunan Provincial Center for Disease Control and Prevention, Changsha 410005, China
2. University of South China, College of Public Health, Hengyang 421001, China
3. NHC Key Laboratory of Medical Virology and Viral Diseases, National Insitute of Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
4. University of South China, College of Public Health, Hengyang 421001, China
- Publication Type:Journal Article
- Keywords:
Human adenovirus-3;
Non-extracting;
Recombinase acid amplification;
Detection
- From:
Chinese Journal of Experimental and Clinical Virology
2019;33(6):653-657
- CountryChina
- Language:Chinese
-
Abstract:
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. The clinical samples of HAdV-3 were detected and compared with the traditional real-time fluorescence quantitative PCR method for nucleic acid extraction.
Results:The sensitivity of the real-time fluorescence RAA method was as high as that of qPCR in the detection of 10 series diluted HAdV-3 strains. The highest corresponding CT value of qPCR was 36.87. The sensitivity of the real-time fluorescence RAA method was similar to that of the real-time fluorescence quantitative PCR method . There was no cross-reaction to other common types of respiratory viruses. The two method were used to detect 56 clinical samples at the same time, and the result were completely consistent.
Conclusions:We provide the first report of the real-time fluorescent RAA assays for the detection of HAdV-3 without extracting nucleic acid and it has high sensitivity and specificity. Is suitable for rapid detection of HAdV-3 in clinical laboratories and on-site unite.
