Multiplex-PCR Method for Species Identification of Coagulase-Positive Staphylococci
Multiplex-PCR Method for Species Identification of Coagulase-Positive Staphylococci
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DOI：
10.1128/jcm.01232-09
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发表时间：
2010-03-01
影响因子：
9.4
通讯作者：
Hiramatsu, Keiichi
中科院分区：
文献类型：
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作者：
Sasaki, Takashi;Tsubakishita, Sae;Hiramatsu, Keiichi
在兽医学中，凝固酶阳性葡萄球菌（CoPS）而不是金黄色葡萄球菌（Staphylococcus aureus）经常被误认为是S。金黄色葡萄球菌菌株，因为它们具有几个共同的表型性状。在兽医临床实验室中，还没有可靠的方法来区分CoPS物种。在本研究中，我们测序的葡萄球菌属的热单核酸酶（nuc）基因，并设计了一个多重PCR（M-PCR）的方法来鉴定种CoPS的nuc基因位点为目标。为了评估灵敏度和特异性，我们使用这种M-PCR方法对374株葡萄球菌菌株进行了检测，这些菌株之前已通过hsp 60测序方法在物种水平上进行了鉴定。我们可以成功地区分S。金黄色葡萄球菌、hyicus、S. schleiferi，S.中间S. pseudintermedius和S. Delphini类群A和B。本方法灵敏度99.8%，特异性100%。我们的M-PCR检测将允许从各种动物种属中常规鉴定CoPS分离株，用于临床兽医诊断。
In veterinary medicine, coagulase-positive staphylococci (CoPS) other than Staphylococcus aureus have frequently been misidentified as being S. aureus strains, as they have several phenotypic traits in common. There has been no reliable method to distinguish among CoPS species in veterinary clinical laboratories. In the present study, we sequenced the thermonuclease (nuc) genes of staphylococcal species and devised a multiplex-PCR (M-PCR) method for species identification of CoPS by targeting the nuc gene locus. To evaluate sensitivity and specificity, we used this M-PCR method on 374 staphylococcal strains that had been previously identified to the species level by an hsp60 sequencing approach. We could successfully distinguish between S. aureus, S. hyicus, S. schleiferi, S. intermedius, S. pseudintermedius, and S. delphini groups A and B. The present method was both sensitive (99.8%) and specific (100%). Our M-PCR assay will allow the routine species identification of CoPS isolates from various animal species for clinical veterinary diagnosis.