猪萨佩罗病毒聚合酶链式反应检测方法的建立及应用  

Establishment and application of a polymerase chain reaction detection method for porcine sapelovirus

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作  者:张伟[1] 韩寅卓 姜永厚[1] ZHANG Wei;HAN Yinzhuo;JIANG Yonghou(College of Life Sciences and Medicine,Zhejiang Sci-Tech University,Hangzhou 310018,China)

机构地区:[1]浙江理工大学生命科学与医药学院,杭州310018

出  处:《浙江理工大学学报(自然科学版)》2024年第5期718-724,共7页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基  金:浙江省自然科学基金项目(LGC21C180002)。

摘  要:猪萨佩罗病毒(Porcine sapelovirus,PSV)是引发猪腹泻的重要病毒之一。为开发一种能快速、准确检测PSV的聚合酶链式反应检测方法,针对PSV的保守区3D基因设计了一对特异性引物,优化了聚合酶链式反应退火温度,分析聚合酶链式反应检测方法特异性和灵敏度,并应用于猪场样品检测。结果表明:建立的聚合酶链式反应检测方法可特异性扩增出PSV目的片段,对其他非靶标猪腹泻病毒则无条带产生,检测低限为5拷贝/μL;在浙江地区采集的88个健康猪粪便样本中检测PSV阳性率为7.95%。该PSV聚合酶链式反应检测方法特异性强、灵敏度高,可应用于PSV前期诊断和防控。Porcine sapelovirus(PSV)is one of the main viruses that cause diarrhea in pig herds.To establish a fast and accurate polymerase chain reaction(PCR)detection method for PSV,specific primers were designed targeting the 3D conserved region sequence of PSV,and the PCR annealing temperature was optimized.The specificity and sensitivity of the established PCR detection method were analyzed and applied to field samples on pig farms.According to the results,the established PCR detection method could specifically amplify the PSV target virus,while it produced no bands for other non-target pig diarrhea viruses,and the lower detection limit was 5 copies/μL.The positive rate of PSV was 7.95%in 88 healthy pig fecal samples collected in Zhejiang province.This PSV PCR detection method demonstrates excellent specificity and sensitivity,and can be applied in the early diagnosis and prevention of PSV.

关 键 词:猪萨佩罗病毒 3D基因 聚合酶链式反应 特异性 灵敏度 阳性率 

分 类 号:TS195.644[轻工技术与工程—纺织化学与染整工程]

 

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