Establishment and verification of real-time fluorescent quantitative PCR detection system for ring virus 6
10.3760/cma.j.issn.1003-9279.2019.06.019
- VernacularTitle: 指环病毒6实时荧光定量PCR检测体系的建立与验证
- Author:
Zhiqiang XIA
1
;
Jun SONG
1
;
Mi LIU
1
;
Qinqin SONG
1
;
Yijin LIU
2
;
Xinhao HAO
2
;
Jun HAN
1
Author Information
1. State Key Laboratory of Infectious Disease Prevention and Control, National Institute of Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
2. Department of Biomedical Engineering, Chengde Medical College, Chengde 067000, China
- Publication Type:Journal Article
- Keywords:
Torque teno virus;
Real-time fluorescent quantitative PCR;
Sensitivity;
Specificity
- From:
Chinese Journal of Experimental and Clinical Virology
2019;33(6):650-652
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a real-time quantitative PCR detection system for Torque teno virus (TTV) and verify the sensitivity and specificity of the detection system.
Methods:Primers and FAM-Eclipse probes were designed based on the TTV6 gene sequence registered in GenBank, and were to establish a real-time fluorescent quantitative PCR detecting way based on the FAM-Eclipse probe, the standard curve was constructed and sensitivity and specificity were analyzed.
Results:A quantitative PCR method for the specific detection of TTV6 were established that the standard curve equation was y=-3.0921x + 28.36, and the amplification efficiency and R2 were 99.6% and 1.000, respectively. The sensitivity of TTV6 was 1.0×10 copies/μl, and there was no cross-reactivity with other viruses. There was 1 case positive for TTV6 out of 56 throat swab samples from the patients with clinical respiratory infection.
Conclusions:The real-time fluorescent quantitative PCR for detecting TTV6 established by FAM-Eclipse probe had the advantages of high sensitivity and specificity. It provides an effective way for detection and quantification of viral content of TTV6 in clinical specimens.
