使用多重PCR检测奶牛乳房炎乳汁中的病原菌  

Use of Multiplex PCR to Detect Pathogens in Milk of Dairy Cows with Mastitis

作　　者：张玉龙 左之才[1] 王宇[1] 崔耀成 李志强 姚彩霞 黄赞恒 才冬杰 ZHANG Yu-long;ZUO Zhi-cai;WANG Yu;CUI Yao-cheng;LI Zhi-qiang;YAO Cai-xia;HUANG Zan-heng;CAI Dong-jie(Key Laboratory of Animal Disease and Human Health of Sichuan Province,College of Veterinary Medicine,Sichuan Agricultural University,Sichuan 625014,China)

机构地区：[1]四川农业大学动物医学院,环境公害与动物疾病四川省高校重点实验室,动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《中国兽医杂志》2021年第11期38-43,共6页Chinese Journal of Veterinary Medicine

基　　金：国家重点研发计划项目(2018YFD0501800);四川省科技计划项目(2018NZ0002);国家现代农业产业技术体系四川肉牛创新团队项目(SCCXTD-2020-13)。

摘　　要：本试验运用多重PCR检测技术检测奶牛乳房炎乳汁中的金黄色葡萄球菌(Staphylococcus aureus)、无乳链球菌(Streptococcus agalactiae)和绿脓杆菌(Pseudomonas aeruginosa)3种病原菌,确认该方法在乳汁病原菌检测中的适用性和有效性,并探讨病原菌的感染情况。选取四川奶牛场送检的191份奶样(经过加州乳房炎检测法检测,其中16份为正常奶样,64份为隐性乳房炎奶样,111份为临床型乳房炎奶样),分别进行多重PCR法和传统细菌学培养法检测,并对照和分析2种方法的差异。结果显示,多重PCR检测法检出135株致病菌,检出率为70.7%;传统细菌学培养法检出99株致病菌,检出率为51.8%;通过卡方分析证明2种检测方法的检出率差异显著(P<0.05)。鉴于多重PCR法表现出较强的特异性、灵敏度,且操作方便,非常适用于3种混合感染病原菌在奶牛临床型乳房炎、隐性乳房炎奶样的检测和流行病学调查。This study compared multiplex PCR method to traditional bacteriological culture method for detection of Staphylococcus aureus、Streptococcus agalactiae and Pseudomonas aeruginosa in milk of dairy cows with mastitis in terms of efficiency and applicability. A total of 191 milk samples(according to the California mastitis test, 16 samples were normal, 64 samples were silent mastitis, and 111 samples were clinical mastitis) from Sichuan dairy farms were tested by multiplex PCR method and traditional bacteriological culture method, and their results were compared and analyzed. The results showed that the multiple PCR method detected 135 of 191 samples, with detection rate of 70.7%, while the traditional bacteriological culture method detected 99 of 191 samples, with detection rate of 51.8%. Use of chi-square test confirmed that the difference in the detection rates between the two detection methods was statistically significant(P<0.05). Given its high specificity and sensitivity, the multiplex PCR method is suitable for the detection and epidemiological investigation of mixed infection pathogens in dairy cows with clinical mastitis and subclinical mastitis.

关 键 词：奶牛乳房炎 病原菌 传统细菌学培养法 多重PCR法 

分 类 号：S854.34[农业科学—临床兽医学]