Establishment and preliminary application of multiplex nano-PCR detection meth-od for Senecavirus A and vesicular stomatitis virus
10.16303/j.cnki.1005-4545.2025.05.07
- VernacularTitle:A型塞内卡病毒和水泡性口炎病毒多重纳米PCR方法的建立及初步应用
- Author:
Xiaojun LI
1
;
Yungang LAN
;
Yue ZHAO
;
Sirui LI
;
Liyuan SHANG
;
Hanrong HUYAN
;
Siwei SONG
;
Wenqi HE
;
Fei GAO
;
Gaili WANG
Author Information
1. 吉林大学动物医学学院,吉林长春 130062;黑龙江省鸡西市现代农业示范基地服务中心,黑龙江鸡西 158170
- Publication Type:Journal Article
- Keywords:
Senecavirus A(SVA);
vesicular stomatitis virus(VSV);
multiplex nano-PCR detection method
- From:
Chinese Journal of Veterinary Science
2025;45(5):934-939,970
- CountryChina
- Language:Chinese
-
Abstract:
The SVA and different serotypes of VSV(VSNJV and VSIV)are susceptible to infect pigs and cause blister injuries to the lips and hoof of pigs.The clinical symptoms of diseases caused by these viruses are very similar,which is easy to cause misdiagnosis.Therefore,a multiplex nano-PCR method was developed for the simultaneous defection of VSV,VSNJV and VSIV.In this stud-y,three pairs of specific primers were designed according to the SVA-P gene,VSNJV-N gene and VSIV-N gene.The optimal annealing temperature and optimal primer concentration were tested,and the reaction system and conditions were optimized.We have developed a novel,rapid and sensitive multiple nano-PCR detection method for simultaneous detection of SVA,VSNJV and VSIV,which was developed by using nano-metal materials.The specific test results showed that the method could specifically amplify the target genes of SVA,VSNJV and VSIV,with no cross-reactivity to PRV,ASFV,PCV2 and PHEV.The sensitivity test results showed that the minimum nucleic acid detection of the method was 10 copies/μL,which sensitivity was great.In addition,the optimal primers showed good reactivity and stability to different batches of enzymes and plasmids.There were 7 among 50 of diseased pig samples were SVA positive by multiple nano-PCR detec-tion method,and 5 out of 50 of diseased pig samples were SVA positive by ordinary single PCR method.Moreover,no VSNJV and VSIV were detected by the two methods.In conclusion,this es-tablished multiple nano-PCR detection method has higher specificity and sensitivity in the detec-tion of SVA,VSNJV and VSIV.And this study could provide technical support for the rapid differ-ential diagnosis,prevention and control of swine viral vesicular diseases in clinical settings.
