摘要
为了建立可快速检测猫杯状病毒(FCV)的方法,本试验采用肽核酸(PNA)设计分子信标荧光探针,优化反应体系和条件,建立了FCV实时荧光定量逆转录PCR(RT-qPCR)检测方法,对该方法的敏感性、特异性和重复性进行验证,并进行临床样本检测。结果显示,建立的RT-qPCR检测FCV参考株具有良好的线性关系(R^(2)=0.9995),最低检测限为1×10^(2)TCID_(50)/mL,对其他相关病毒无有效响应,组内变异系数和组间变异系数均小于1%。应用该方法检测33份临床样本,阳性率为51.5%。由此可见,本试验建立的RT-qPCR敏感性、特异性和重复性均良好,可为FCV的早期快速诊断和疫病防控等提供检测手段。
To develop a method for rapid detection of feline calicivirus(FCV),this study designed a peptide nucleic acid(PNA)molecular beacon fluorescent probe,optimized the reaction system and conditions,and established a real-time quantitative reverse transcription PCR(RT-qPCR)detection method for FCV.The sensitivity,specificity,and repeatability of this method were validated,and clinical samples were tested.The results showed that the established RT-qPCR had a good linear relationship for the FCV reference strain(R^(2)=0.9995),with a minimum detection limit of 1×10^(2)TCID_(50)/mL.There was no effective response to other related viruses,and both intra-assay and inter-assay coefficients of variation were below 1%.When applied to 33 clinical samples,the positive detection rate was 51.5%.These findings indicate that the RT-qPCR method established in this study is highly sensitive,specific,and repeatable,providing a reliable tool for early and rapid diagnosis of FCV and aiding in disease prevention and control.
作者
李静怡
熊雷
黄莉璎
刘晓鹏
杨盛奋
金京勋
王弋
LI Jingyi;XIONG Lei;HUANG Liying;LIU Xiaopeng;YANG Shengfen;JIN Jingxun;WANG Yi(Dongguan Bioshine Biotechnology Co.,Ltd.,Dongguan 523808,China;College of Pharmacy,Jinan University,Guangzhou 510632,China;Guangzhou Yuanbo Medical Technology Co.,Ltd.,Guangzhou 510700,China)
出处
《中国兽医杂志》
CAS
北大核心
2024年第12期64-72,共9页
Chinese Journal of Veterinary Medicine
基金
东莞市引进创新创业领军人才计划资助(2017-16)
广州开发区创新创业领军人才(2021-L010)
暨南大学-东莞博盛生物科技有限公司的博士后基金(BS-XM2024001)。
