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当前位置: 首页 > 文章 > 奶牛源金黄色葡萄球菌新疆流行株生物被膜形成及相关基因的分布与转录水平 南方农业学报 2019,50 (8) 1829-1835
Position: Home > Articles > Analysis of biofilm formation,related genes distribution and transcription level in Xinjiang isolates of Staphylococcus aureus from cows Journal of Southern Agriculture 2019,50 (8) 1829-1835

奶牛源金黄色葡萄球菌新疆流行株生物被膜形成及相关基因的分布与转录水平

作  者：
伍晔晖;孟庆玲;乔军;李静;蔡扩军;王登峰;才学鹏
单  位：
新疆维吾尔自治区畜牧科学院兽医研究所;石河子大学科技学院;乌鲁木齐市动物疾病控制与诊断中心;中国农业科学院兰州兽医研究所
关键词：
金黄色葡萄球菌;生物被膜;黏附素;转录水平;新疆
摘  要：
【目的】了解金黄色葡萄球菌(Staphylococcus aureus,SA)临床分离株的生物被膜形成情况及黏附素和ica操纵子与生物被膜形成能力的相关性,为系统掌握新疆地区奶牛源SA流行株的分子特征及有效防治奶牛乳房炎提供科学依据。【方法】收集临床分离鉴定的164株SA新疆流行株,采用结晶紫半定量黏附试验(MPA)测定各流行株的体外生物被膜形成能力,同时通过PCR和实时荧光定量PCR(qRT-PCR)对SA生物被膜形成相关基因转录水平进行检测分析。【结果】164株奶牛源SA新疆流行株的生物被膜阳性率为86.0%(141株),其中弱(+)生物被膜形成有69株(占42.1%)、中等(++)生物被膜形成有38株(占23.2%)、强(+++)生物被膜形成有34株(20.7%)。在141株MPA阳性SA分离株中,clfA、clfB、fnbA、fnbB、cna、fib、icaA、icaC和icaD基因检出率分别为83.7%(n=118)、58.9%(n=83)、75.2%(n=106)、78.7%(n=111)、75.9%(n=107)、58.9%(n=83)、90.1%(n=127)、79.4%(n=112)和100.0%(n=141);ica基因(icaA、icaC和icaD)检出率总体上高于其他生物被膜形成相关基因,且生物被膜形成能力强(+++和++)的菌株尤为明显;clfB、fnbA、cna和fib基因检出率则表现为MPA阴性菌株高于MPA阳性菌株。9个生物被膜形成相关基因中有7个基因(clfA、fnbA、fnbB、fib、cna、icaC和icaD)在强(+++)生物被膜形成能力SA分离株中的表达量显著上调(P<0.05)。【结论】奶牛源SA新疆流行株生物被膜形成率高,生物被膜形成相关基因携带率也较高,其中clfA、fnbA、fnbB、fib、cna、icaC和icaD基因的表达与SA生物被膜形成密切相关。
译  名：
Analysis of biofilm formation,related genes distribution and transcription level in Xinjiang isolates of Staphylococcus aureus from cows
作  者：
WU Ye-hui;MENG Qing-ling;QIAO Jun;LI Jing;CAI Kuo-jun;WANG Deng-feng;CAI Xue-peng;College of Animal Science and Technology,Shihezi University;Animal Disease Control and Diagnosis Center in Urumqi;Institute of Veterinary Medicine,Xinjiang Academy of Animal Science;Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences;
关键词：
Staphylococcus aureus;;biofilm;;adhesion;;transcription level;;Xinjiang
摘  要：
【Objective】The aim of the study was to detect biofilm formation and correlation between adhesin,ica operon with biofilm formation ability of the clinical isolates of Staphylococcus aureus(SA),which would provide an insight into molecular characteristic of SA strains of dairy cows in Xinjiang and provide a scientific basis for the prevention and treatment of dairy cow mastitis.【Method】The 164 strains of SA strains in Xinjiang from clinical isolates were collected. The in vitro biofilm formation ability of the bacteria were then evaluated by a microtiter plate assay(MPA),and the biofilm-associated genes were detected by PCR and real-time fluorescence quantitative PCR(qRT-PCR).【Result】The positive rate of biofilm formation was 86.0%(141 isolates)among the 164 SA isolates in Xinjiang from cows. Of the 141 biofilm positive isolates,69(42.1 %)were weak(+)biofilm formation,38(23.2%)were medium(++)biofilm formation and 34(20.7%)were strong(+++)biofilm formation,respectively. Among 141 MPA-positive SA isolates,the detection rates of biofilm-associated genes clfA,clfB,fnbA,fnbB,cna,fib,icaA,icaC and icaD were 83.7%(n=118),58.9%(n=83),75.2%(n=106),78.7%(n=111),75.9%(n=107),58.9%(n=83),90.1%(n=127),79.4%(n=112)and 100.0%(n=141),respectively. The detection rates of ica genes(icaA,icaC and icaD)were higher than those of other biofilm-associated genes,and the strains with strong biofilm formation ability(+++ and ++)were particularly obvious. The detection rate of clfB,fnbA,cna and fib genes was higher in MPA-negative strains than in MPA-positive strains. Among the nine bbiofilm-forming related genes,seven biofilm-forming genes(clfA,fnbA,fnbB,fib,cna,icaC,and icaD)were significantly up-regulated in the SA isolates with strong(+++)biofilm-forming ability(P<0.05).【Conclusion】The formation rate of SA biofilm from dairy cows in Xinjiang is high,and the biofilm formation-related genes have a high carrying rate.The expression of clfA,fnbA,fnbB,fib,cna,icaC and icaD genes is closely related to biofilm formation.

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