Establishment of TaqMan RT-PCR assay for Wuxiang virus
10.3760/cma.j.cn112866-20220310-00056
- VernacularTitle:武乡病毒TaqMan RT-PCR检测方法的建立
- Author:
Danhe HU
1
;
Xiaohui YAO
;
Shihong FU
;
Fan LI
;
Tianmeng GU
;
Junjie CHEN
;
Ying HE
;
Jiayu YIN
;
Songtao XU
;
Xiangdong LI
;
Kai NIE
;
Huanyu WANG
;
Guodong LIANG
Author Information
1. 扬州大学兽医学院，扬州 225009
- Keywords:
Wuxiang virus;
TaqMan RT-PCR;
Detection
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(4):460-464
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a real-time fluorescent quantitative TaqMan reverse transcription polymerase chain reaction (Real-time RT-PCR) detection method for Wuxiang virus (WUXV).Methods:All gene sequences of WUXV were downloaded from GenBank, and multi-sequence alignment analysis was performed using Mega-X. Primers and probes designed for the highly conservative region of S-segment genes were selected to evaluate the specificity, sensitivity and stability of detection reactions.Results:The established method can specifically detect WUXV and does not cross-react with various arboviruses. The lowest detection limit was 1 pfu/ml. The inter-batch variation coefficients of repeated detection cycle threshold ( Ct) of the same sample were all less than 1.00%. TaqMan RT-PCR was used to detect 30 batches of sandflies nucleic acid samples, and 7 of them showed positive amplification curve of WUXV. Conclusions:TaqMan RT-PCR with high sensitivity, specificity and repeatability has been successfully established, which can be used to screen large quantities of samples of WUXV.
