Establishment of real-time fluorescent quantitative PCR for detection of torque teno virus types 7, 8 and 10
10.3760/cma.j.cn112309-20200921-00444
- VernacularTitle:指环病毒7型、8型与10型实时荧光定量PCR检测体系的建立
- Author:
Zhiqiang XIA
;
Juan SONG
;
Dong XIA
;
Qinqin SONG
;
Wenjun WANG
;
Ruifang WANG
;
Bingtian SHI
;
Mi LIU
;
Geng HU
;
Yanhai WANG
;
Jun HAN
- From:
Chinese Journal of Microbiology and Immunology
2021;41(3):190-194
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.
