马腺疫链球菌fneB基因LAMP检测方法的建立及应用  

Development and Application of a LAMP Method for Detection of fneB Gene of Streptococcus equi

作　　者：买占海 沙娅·奴尔兰 王德超 张敖云图雅 侯宇[1] 况玲[1] MAI Zhanhai;SHA Ya·Nuerlan;WANG Dechao;ZHANG Aoyuntuya;HOU Yu;KUANG Ling(Xinjiang Agricultural University,Urumqi 830052,China;Ili Prefecture Animal Disease Prevention and Control Center,Ili 835800,China)

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052 [2]伊犁州动物疾病预防与控制中心,伊宁835800

出　　处：《中国动物传染病学报》2022年第1期106-111,共6页Chinese Journal of Animal Infectious Diseases

基　　金：自治区普通高等学校教学改革研究项目(2017JG042);新疆农业大学研究生科研创新项目(XJAUGRI2017-034)。

摘　　要：为了建立快速简便的fneB-LAMP检测法用于马腺疫病原马链球菌马亚种(S.equi)的检测,本试验采用恒温水浴进行LAMP扩增,建立可视化fneB-LAMP检测方法。结果显示:对大肠埃希菌、无乳链球菌、金黄色葡萄球、蜡状芽孢杆菌和克雷伯杆菌5种菌与标准S.equi菌株检测,只能够检测出标准阳性菌株;对新疆昭苏县3个马场临床检查疑似马腺疫的鼻拭子和脓汁共40份样品进行fneB-LAMP检测,其中有7株分离菌呈阳性;与普通PCR检测方法相比,LAMP检测方法灵敏度为8×10^(-6)μg/μL,较普通PCR方法高;该方法在粗提模板的情况下只需简单的恒温仪器反应1 h可得到可视化的检测结果。本试验成功建立了马腺疫病原靶向fneB基因LAMP检测的方法,能够快速准确的检测出S.equi,为今后快速诊断马腺疫疾病提供参考。The objective of the present study was to develop a simple LAMP method to detect the fneB gene of Streptococcus equi subspecies equi.In this method,the target sequence of the fneB gene was amplifi ed in a constant temperature bath.The preliminary visual fneB-LAMP detection method was tested for its specifi city by comparing standard S.equi strains with 5 strains of Escherichia coli,Streptococcus lactis,Staphylococcus aureus,Bacillus cereus and Klebsiella.The results showed that standard positive S.equi strains could be detected in this method.Forty nasal swab and pus samples of suspected of equine adenovirus were collected from 3 horse farms in Zhaosu county,Xinjiang and 7 samples were positive by fneB-LAMP method.In addition,the sensitivity of the LAMP detection was higher than common PCR.This method could obtain visual detection results by a simple constant temperature instrument by amplifi cation for 1 h after rough extraction of the template.In conclusion,a LAMP method developed here could be used for quickly and accurately detecting fne B gene of S.equi,which be developed a rapid diagnosis of strangle.

关 键 词：马腺疫 环介导等温扩增技术 fneB基因 马链球菌马亚种 

分 类 号：S852.611[农业科学—基础兽医学]