对五种qPCR和PCR方法在弓形虫诊断中的比较研究  

A comparative study of five qPCR and PCR methods for Toxoplasma gondii diagnosis

作　　者：黄艳丽 陈亚娜 白邵缘 周国庆 崔灿 李宇渊 黄思扬 原霖 HUANG Yanli;CHEN Ya'na;BAI Shaoyuan;ZHOU Guoqing;CUI Can;LI Yuyuan;HUANG Siyang;YUAN Lin(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Beijing Zhongke Gene Technology Co.,Ltd.,Beijing 102600,China;Pudao(Beijing)Standard Technology Co.,Ltd.,Beijing 102600,China;Joint International Research Laboratory of A griculture and A gri-Product Safety,Ministry of Education,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]北京中科基因技术股份有限公司,北京102600 [3]普道(北京)标准技术有限公司,北京102600 [4]教育部农业与农产品安全国际合作联合实验室,江苏扬州225009

出　　处：《中国兽医科学》2025年第3期346-353,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2022YFC2304003);江苏省“双创计划”-双创人才项目。

摘　　要：为了评估国内外4个标准中涉及的5个PCR和实时定量PCR(qPCR)方法在弓形虫病诊断中的性能,本研究基于弓形虫速殖子评估了各检测方法的敏感性、重复性和特异性,并通过检测62份猫粪便样品对上述诊断方法进行了比较分析。结果显示,所有标准方法都有较好的特异性和重复性,其中检验检疫标准中的qPCR方法、WOAH标准的qPCR方法和农业标准的PCR方法最低检测限最小(速殖子的浓度约为每微升0.375个),其次是国家标准的PCR方法(速殖子的浓度约为每微升3.75个),检验检疫标准的PCR方法敏感性最低(速殖子的浓度约为每微升375个)。对62份猫粪便样品进行检测分析发现,敏感性最高(99.23%)的方法为检验检疫标准的qPCR方法,特异性最高(99.13%)的是国家标准的PCR方法,敏感性最低(91.73%)的是检验检疫标准中的PCR方法,特异性最低(96.82%)的是农业标准的PCR方法。本研究为弓形虫病诊断标准方法的选择提供了参考。To evaluate the performance of 5 PCR and real-time quantitative PCR(q PCR)methods involved in 4 domestic and international standards for diagnosing toxoplasmosis,the sensitivity,repeatability,and specificity of each detection method were assessed based on Toxoplasma gondii tachyzoites.Sixty-two cat fecal samples were used to compare the diagnostic methods mentioned above.The results showed that all standard methods have good specificity and repeatability.Among them,the q PCR method in the SN/T,the q PCR method in the WOAH,and the PCR method in the NY/T had the lowest detection limit(approximately0.375 tachyzoites/μL),followed by the PCR method in the GB/T(approximately 3.75 tachyzoites per microlitre).The PCR method in the SN/T has the lowest sensitivity(approximately 375 tachyzoites per microlitre).Analysis of testing 62 cat fecal samples revealed that the q PCR method in the SN/T had the highest sensitivity(99.23%),the PCR method in the GB/T had the highest specificity(99.13%),the PCR method in the SN/T had the lowest sensitivity(91.73%),and the PCR method in the NY/T had the lowest specificity(96.82%).This study provides a reference for the selection of standard methods for diagnosing toxoplasmosis。

关 键 词：弓形虫 标准 PCR qPCR 方法评价 

分 类 号：S852.723[农业科学—基础兽医学]