鸡圆圈病毒3型SYBR Green Ⅰ实时荧光定量PCR方法的建立与应用OACSTPCD
Establishment and application of SYBR Green Ⅰ real-time fluorescence quantitative PCR method for chicken circle virus type 3
本研究针对圆圈病毒3型(GyV3)VP2基因构建标准质粒,以此作为模板建立标准曲线,在优化qPCR反应体系的基础上,对该方法的特异性、敏感性和重复性进行评估.结果显示,该方法仅对GyV3特异性扩增,最低检测模板浓度为1.585×101copies/μL,组内和组间重复性试验的变异系数均小于1.0％.用本研究建立的qPCR方法对50份临床样品进行检测,阳性检出率为10％(5/50),高于常规PCR的阳性检出率8％(4/50).上述结果表明,本研究建立的实时荧光定量PCR方法特异性强、重复性好、灵敏度高,可用于临床快速检测GyV3.
In this study,a standard plasmid based on GyV3 VP2 gene was constructed and used as a template to establish a standard curve.On the basis of optimizing the qPCR reaction system,the specificity,sensitivity and repeatability of this method were evaluated.The results revealed the method was only specific for GyV3 amplification,the minimum detection template concentration was 1.585 × 101 copies/μL,and the coefficient of variation in both intra-class and intra-group repea-tability tests was less than 1.0％.The qPCR method established in this study was used to detect 50 clinical samples,and the detection rate was 10％(5/50),higher than the conventional PCR detection rate of 8％(4/50).The results above indicate that the real-time fluorescent quantitative PCR method established in this study has high specificity,reproducibility,and sensitivity and can be utilized for quick clinical detection of GyV3.
牛群;庄丽云;雷天宇;戴婷婷;孟庆福;刘荣昌;何华;郑新添
龙岩学院预防兽医学与生物技术福建省高等学校重点实验室,福建龙岩 364000||河南农业大学动物医学院,河南郑州 450046龙岩学院预防兽医学与生物技术福建省高等学校重点实验室,福建龙岩 364000||福建农林大学动物科学学院,福建福州 350000龙岩学院预防兽医学与生物技术福建省高等学校重点实验室,福建龙岩 364000福建省农业科学院畜牧兽医研究所,福建福州 350013||福建省禽病防治重点实验室,福建福州 350013河南农业大学动物医学院,河南郑州 450046
畜牧业
圆圈病毒3型VP2基因实时荧光定量PCR
gyrovirus 3VP2 genereal-time fluorescence quantitative PCR
《中国兽医科学》 2024 (001)
56-62 / 7
国家现代农业产业技术体系资助项目(CARS-42);龙岩学院预防兽医学与生物技术福建省高等学校重点实验室开放基金项目(2022KF03);河南农业大学"拔尖人才"项目(30500737)
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