Rapid Identification of Staphylococcal Infection in Dairy Goat Raw Milk by a Multiplex PCR Assay

應用多引子PCR快速鑑定山羊生乳之葡萄球菌感染

朱紀實(Chi-Shih Chu) ； 賴治民(Jyh-Mirn Lai) ； 余章游(Chang-You Yu) ； 張文興(Wing-Shin Chang) ； 林欣虹(Hsing-Hong Lin) ； 蘇耀期(Yao-Chi Su)

《臺灣獸醫學雜誌》 36卷3期 (2010/09) Pp. 182-190

https://doi.org/10.7009/TVJ.201009.0182

摘要

為改善乳用山羊乳房炎之診斷，本研究發展一套多引子PCR技術作為針對葡萄球菌屬感染快速鑑定之診斷工具，設計依據葡萄球菌屬特有的16S rRNA基因之引子對檢測葡萄球菌屬菌種，而排除其他細菌菌種，並增幅金黃色葡萄球菌特有之nuc基因之引子對來區分S. aureus和非金黃色葡萄球菌之葡萄球菌屬細菌，同時依據是否增幅出mecA基因之PCR產物，作為受檢菌株是否具有methicillin抗藥性之指標（如methicillin resistant staphylococcus aureus; MRSA）。總計收集3,427個次臨床乳羊場生乳樣本，配合傳統培養、API生化套組及PCR方法進行鑑定，結果顯示41.4%的細菌感染，優勢菌種分別為E-coli、Coagulase-negative staphylococci和S. aureus，這些菌種於各場間之盛行率存在著差異。S. aureus除A、B、D、E場之外，各場間皆為單一基因型(genotype A1)。

關鍵字

多引子聚合酶鏈反應 ； 乳山羊 ； 葡萄球菌

並列摘要

To improve the diagnosis accuracy of staphylococcal infection in dairy goat with mastitis, we develops a multiplex polymerase chain reaction (PCR) assay by targeting specific regions of staphylococcal 16S rRNA for identification of staphylococcal infection, using specific nuc gene of S. aureus to distinguish S. aureus and the coagulase negative Staphylococci (CNS), and mecA gene responsible for oxacillin or methicillin resistance to determine methicillin resistant staphylococcus aureus (MRSA). The Analytical Profile Index (API) method, and PCR method were used to examine 3,427 milk samples from subclinically infected herds with traditional culture, and revealed 41.4% bacterial infection. E. coli, coagulase-negative staphylococci and S. aureus were the predominant bacteria and their prevalence differed among farms. Genotype Al S. aureus was clonally disseminated on most farms, except farms A, B, D, and E.

並列關鍵字

dairy goat ； multiplex PCR ； Staphylococcus