Development and application of a triplex TaqMan fluorescent quantitative PCR assay for simultaneous detection of Senecavirus A,foot-and-mouth disease virus and porcine teschovirus
10.16303/j.cnki.1005-4545.2025.01.04
- VernacularTitle:A型塞内卡病毒、口蹄疫病毒和猪捷申病毒3重TaqMan荧光定量PCR检测方法的建立
- Author:
Shiqi GAN
1
;
Qianhe WEI
;
Yuchen NI
;
Jianbo NI
;
Xiuling ZHAO
;
Wanyu DONG
;
Yings-han ZHOU
;
Xiaodu WANG
Author Information
1. 浙江农林大学动物科技学院·动物医学院浙江省畜禽绿色生态健康养殖应用技术研究重点实验室/动物健康互联网检测技术浙江省工程实验室/浙江省动物医学与健康管理国际科技合作基地/中澳动物健康大数据分析联合实验室,浙江 杭州 311300
- Publication Type:Journal Article
- Keywords:
Senecavirus A;
foot-and-mouth disease virus;
porcine teschovirus;
fluorescent quantita-tive RT-PCR
- From:
Chinese Journal of Veterinary Science
2025;45(1):22-29
- CountryChina
- Language:Chinese
-
Abstract:
Primers and probes were designed based on the conserved regions of Senecavirus A(SVA),foot-and-mouth disease virus(FMDV),and porcine teschovirus(PTV)and used to devel-op a TaqMan fluorescent quantitative PCR method for detecting the above-mentioned three viru-ses.The triplex fluorescent quantitative PCR system was developed using recombinant positive plasmids containing conserved sequences of the three viruses as templates.After optimizing the conditions,the specificity,sensitivity,repeatability,standard curve,and mixed infection model were evaluated,and the constructed triplex fluorescent quantitative PCR was used for clinical detection.The results showed that this method could specifically detect SVA,FMDV and PTV without cross-reactivity with other pathogens with the minimal detection concentrations for SVA,FMDV,and PTV as low as 1X101 copies/μL,respectively.The coefficients of variation within and between groups were less than 5％.Furthermore,none of the three viruses were detected in 126 samples.The above results indicate that this method is highly specific,sensitive,and stable,making it suit-able for clinical detection.
