中国动物检疫2024，Vol.41Issue(8)：105-111,7.DOI:10.3969/j.issn.1005-944X.2024.08.019
牛结核病荧光定量PCR引物探针的组合筛选比较
Comparison of Combinations of Fluorescent Quantitative PCR Primers and Probe for Bovine Tuberculosis
摘要
Abstract
In order to compare the specificity and sensitivity of different combinations of fluorescent quantitative PCR(qPCR)primers and probe for bovine tuberculosis,and to verify their clinical effects,11 combinations proposed by international,national,industrial and local standards and reported by literatures were compared and verified using national secondary standard materials of Mycobacterium bovis genomic DNA,nucleic acids of reference strain as well as clinical milk samples.The results showed that the 11 combinations all had no non-specific amplification,indicating good specificity;the primers originated from national standard IS1081 could reach a minimum detection limit of 1.4×10-1 with the highest sensitivity;the reproducibility of other combinations was good(CV＜10%),except for the national standard primers(Mtb);for 30 clinical samples,the detection rate of the positive samples detected by the primers originated from the national standard IS1081 was significantly higher than that by other combinations.In conclusion,the combination of primers and probe in national standard IS1081 was specific and sensitive,which could receive better clinical effects and was proposed to detect bovine tuberculosis by qPCR in laboratory.关键词
牛结核病/结核分枝杆菌复合群/牛分枝杆菌/荧光定量PCR/引物探针Key words
bovine tuberculosis/Mycobacterium tuberculosis complex/Mycobacterium bovis/fluorescent PCR/primer and probe分类
畜牧业引用本文复制引用
毛迎雪,孙淑芳,胡莉萍,樊晓旭,亓菲,张皓博,曲瑶,南文龙,刘蒙达,苏华彬,姜志强,刘建柱..牛结核病荧光定量PCR引物探针的组合筛选比较[J].中国动物检疫,2024,41(8):105-111,7.基金项目
"十四五"国家重点研发计划项目(2022YFD1800701) （2022YFD1800701）
国家现代农业产业技术体系资助项目(CARS-36) （CARS-36）
