Establishment and application of a TaqMan real-time fluorescence quantitative PCR for detection of tree shrew adenovirus(TAV)
10.3969/j.issn.1671-7856.2018.03.014
- VernacularTitle:树鼩腺病毒TaqMan实时荧光定量PCR检测方法的建立及初步应用
- Author:
Qingkai SONG
1
;
Xiaofei LI
;
Yurun MIAO
;
Zhicheng ZHANG
;
Xuan WANG
;
Yuan YUAN
;
Jiejie DAI
;
Xiaomei SUN
Author Information
1. 中国医学科学院/北京协和医学院医学生物学研究所树鼩种质资源中心
- Keywords:
tree shrew adenovirus;
3'conserved region;
TaqMan;
real-time fluorescence quantitative PCR
- From:
Chinese Journal of Comparative Medicine
2018;28(3):72-77
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a quick and accurate method for detection of tree shrew adenovirus(TAV) using TaqMan real-time fluorescence quantitative PCR. Methods Based on the published TAV genome sequence, a 3' conserved sequence was used to design specific probe primers. A standard curve was prepared using a recombinant plasmid containing the target gene fragment. A real-time fluorescence quantitative PCR method was established for detecting TAV based on TaqMan probe. Results The detection method was specific and was not cross-reactive with other common pathogens. The detection limit of the method was 3.7 copies/μL,showing a high sensitivity. The correlation coefficient was 0.998, and the efficiency was 95.7%. The amplification result showed a fine linear relationship,and the repeatability test effect was good. Conclusions The TAV real-time quantitative PCR detection method based on TaqMan probe has been successfully established. It has high sensitivity and reproducibility and can be applied to early detection of TAV infection.
