Establishment of TaqMan RT-qPCR assay for the detection Getah virus
10.3760/cma.j.cn112866-20200120-00011
- VernacularTitle:盖塔病毒TaqMan RT-qPCR检测方法的建立与评价
- Author:
Tianyuan WU
1
;
Shihong FU
;
Qikai YIN
;
Jierong ZHAO
;
Fan LI
;
Ying HE
;
Songtao XU
;
Guodong LIANG
;
Kai NIE
;
Guang YANG
;
Huanyu WANG
Author Information
1. 暨南大学基础医学与公共卫生学院，广州 510000
- Keywords:
Getah virus;
TaqMan;
RT-qPCR;
Molecular diagnosis
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(2):205-208
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a sensitive and specific real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) method for rapid detection of Getah virus (GETV).Methods:All the gene sequences of GETV were downloaded from GenBank database. Clustal X was used for sequence alignment, and specific primers and probes were designed according to highly conserved regions; we established a standard curve using the nucleic acid of GETV as a standard, and the sensitivity, specificity and stability of this method were evaluated respectively.Results:This method could specifically detect GETV and has no cross-reactivity with multiple arboviruses; the sensitivity was 1.0×10 pfu/ml, and the intra-assay and inter-assay coefficients of variation were less than 1%. One case was GETV positive in 196 batches of mosquitoes collected from Hunan province, Hebei province, Fujian province and Chongqing city.Conclusions:We established a TaqMan probe real-time quantitative RT-PCR with high sensitivity and specificity which can be used for screening.
