畜牧与兽医2025，Vol.57Issue(8)：100-107,8.
鸡毒支原体和鸡滑液囊支原体的双重PCR检测方法建立及其在海南文昌鸡主养区感染情况调查
Establishment of duplex PCR method for detection of Mycoplasma gallisepticum and Mycoplasma synoviae and etiology investigation of Wenchang chicken breeding areas in Hainan Province
摘要
Abstract
This study was to establish a duplex PCR method for detection of Mycoplasma gallisepticum(MG)and Mycoplasma synovium(MS)so as to clarify the infection status and epidemic characteristics of MG and MS in the main Wenchang chicken breeding areas in Hainan Province.According to the conserved sequences of the MG mgC2 gene and the MS vlhA gene,primers were respectively designed.The duplex PCR method for detection of MG and MS was established by optimizing the reaction conditions and procedures.And the sensitivity,specificity and repeatability of the method were verified.At the same time,the established method was compared with the national agricultural industry standard method to verify its accuracy.Finally,1 510 throat swab samples colleted from the main Wenchang chicken breeding areas in Hain-an Province were detected with this method.The results showed that the minimum detection limit of the duplex PCR method for detecting MG and MS positive plasmids was 2.1×102 copies/μL.There was no cross reaction with Newcastle disease,Marek's disease,Infectious bursal disease and Infectious Laryngotracheitis.Compared with the MG and MS single PCR,the positive coincidence rates of the two methods were 100％and 98.83％,respectively.The results of the pathogenic detection showed that the infection rate of MG was 11.79％(95％CI:10.16％-13.42％),that of MS was 25.96％(95％CI:23.75％-28.17％),and the mixed infection rate of MG and MS was 7.42％(95％CI:6.10％-8.74％).MG and MS had a high incidence in spring,most seriously in medium-sized farms,adult chickens and commercial broilers.The present results indicated that the duplex PCR method of MG and MS established in this study had good sensitivity,specificity,stability and accuracy,and could be used in clinical detection.Currently,there are infections of MG and MS in Wenchang chicken breeding areas of Hainan Province,so the prevention and control of MG and MS should be strengthened.关键词
鸡毒支原体/鸡滑液囊支原体/双重PCR/病原学/文昌鸡Key words
Mycoplasma gallisepticum/Mycoplasma synoviae/Duplex PCR/etiology/Wenchang chicken分类
农业科技引用本文复制引用
薛琛璐,张艳,刘海隆,陈素贞,董亚童,晁哲,魏立民,刘圈炜..鸡毒支原体和鸡滑液囊支原体的双重PCR检测方法建立及其在海南文昌鸡主养区感染情况调查[J].畜牧与兽医,2025,57(8):100-107,8.基金项目
海南省农业科学院畜牧兽医研究所所级重点任务项目(HNXM2024ZD01) （HNXM2024ZD01）
农业农村部优势特色产业集群(文昌鸡)项目(WC-SCICP20211106) （文昌鸡）
国家现代农业肉鸡产业技术体系专项(CARS-41-Z12) （CARS-41-Z12）
