Kuo Hung-Chih, Lo Dan-Yuan, Chen Chiou-Lin, Tsai Yun-Long, Ping Jia-Fong, Lee Chien-Hsien, Lee Pei-Yu Alison, Chang Hsiao-Fen Grace
Yunlin-Chiayi-Tainan of Animal Disease Diagnostic Center, Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan.
GeneReach Biotech, Taichung, Taiwan.
Poult Sci. 2017 Jan 1;96(1):35-41. doi: 10.3382/ps/pew228. Epub 2016 Jul 7.
Mycoplasma synoviae (MS), causing respiratory diseases, arthritis, and eggshell apex abnormalities in avian species, is an important pathogen in the poultry industry. Implementation of a biosecurity plan is important in MS infection management. Working on a field-deployable POCKIT™ device, an insulated isothermal polymerase chain reaction (iiPCR) assay has a potential for timely MS detection on the farm. The MS iiPCR assay had limit of detection 95% of about 9 genome equivalents by testing serial dilutions of a standard DNA. The detection endpoint of the assay for detection of MS genomic DNA was comparable to a reference real-time PCR. The assay did not crossreact with other important avian pathogens, including avian reovirus, Mycoplasma gallisepticum, Staphylococcus aureus, Escherichia coli, Pasteurella multocida, and Salmonella Pullorum. When 92 synovial fluid and respiratory tract swab samples collected from chickens, turkeys, and geese suspected of MS infection were tested, the clinical performance of the MS iiPCR had 97.8% agreement (Cohen's kappa value, 0.95) with that of the reference real-time PCR. In conclusion, the MS iiPCR/POCKIT™ system, working with field-deployable manual or automatic nucleic acid extraction methods, has potential to serve as a rapid and sensitive on-site tool to facilitate timely detection of MS.
滑膜支原体（MS）可导致禽类呼吸道疾病、关节炎和蛋壳顶端异常，是家禽业中的一种重要病原体。实施生物安全计划对MS感染管理很重要。基于可现场部署的POCKIT™设备开展工作，一种隔热等温聚合酶链反应（iiPCR）检测方法有潜力在农场及时检测出MS。通过对标准DNA进行系列稀释测试，MS的iiPCR检测方法的检测限约为9个基因组当量，95%置信度。该检测方法检测MS基因组DNA的检测终点与参考实时荧光定量PCR相当。该检测方法不与其他重要的禽类病原体发生交叉反应，包括禽呼肠孤病毒、鸡毒支原体、金黄色葡萄球菌、大肠杆菌、多杀性巴氏杆菌和鸡白痢沙门氏菌。当对从疑似感染MS的鸡、火鸡和鹅采集的92份滑液和呼吸道拭子样本进行检测时，MS的iiPCR的临床性能与参考实时荧光定量PCR的一致性为97.8%（科恩kappa值，0.95）。总之，MS的iiPCR/POCKIT™系统与可现场部署的手动或自动核酸提取方法配合使用，有潜力作为一种快速、灵敏的现场工具，便于及时检测出MS。
