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调查在爱尔兰兽医医院从伴侣动物中分离的葡萄球菌属种时,常规使用的表型方法与基因型方法的效果比较。

Investigations on the efficacy of routinely used phenotypic methods compared to genotypic approaches for the identification of staphylococcal species isolated from companion animals in Irish veterinary hospitals.

机构信息

Biosciences Research Institute, Athlone Institute of Technology, Dublin Rd., Athlone, Co. Westmeath, Ireland ; Department of Life and Physical Science, School of Science, Athlone Institute of Technology, Dublin Rd., Athlone, Ireland.

Biosciences Research Institute, Athlone Institute of Technology, Dublin Rd., Athlone, Co. Westmeath, Ireland.

出版信息

Ir Vet J. 2013 May 1;66(1):7. doi: 10.1186/2046-0481-66-7. eCollection 2013.

Abstract

BACKGROUND

Identification of Staphylococci to species level in veterinary microbiology is important to inform therapeutic intervention and management. We report on the efficacy of three routinely used commercial phenotypic methods for staphylococcal species identification, namely API Staph 32 (bioMérieux), RapID (Remel) and Staph-Zym (Rosco Diagnostica) compared to genotyping as a reference method to identify 52 staphylococcal clinical isolates (23 coagulase positive; 29 coagulase negative) from companion animals in Irish veterinary hospitals.

RESULTS

Genotyping of a 412 bp fragment of the staphylococcal tuf gene and coagulase testing were carried out on all 52 veterinary samples along with 7 reference strains. In addition, genotyping of the staphylococcal rpoB gene, as well as PCR-RFLP of the pta gene, were performed to definitively identify members of the Staphylococcus intermedius group (SIG). The API Staph 32 correctly identified all S. aureus isolates (11/11), 83% (10/12) of the SIG species, and 66% (19/29) of the coagulase negative species. RapID and Staph-Zym correctly identified 61% (14/23) and 0% (0/23) respectively of the coagulase-positives, and 10% (3/29) and 3% (1/29) respectively of the coagulase-negative species.

CONCLUSIONS

Commercially available phenotypic species identification tests are inadequate for the correct identification of both coagulase negative and coagulase positive staphylococcal species from companion animals. Genotyping using the tuf gene sequence is superior to phenotyping for identification of staphylococcal species of animal origin. However, use of PCR-RFLP of pta gene or rpoB sequencing is recommended as a confirmatory method for discriminating between SIG isolates.

摘要

背景

在兽医微生物学中,鉴定葡萄球菌至种水平对于指导治疗干预和管理非常重要。我们报告了三种常用的商业表型方法在鉴定葡萄球菌种方面的功效,即 API Staph 32(生物梅里埃)、RapID(雷美)和 Staph-Zym(Rosco Diagnostica),并将其与作为参考方法的基因分型进行比较,以鉴定来自爱尔兰兽医医院的 52 株临床分离的葡萄球菌(23 株凝固酶阳性;29 株凝固酶阴性)。

结果

对所有 52 个兽医样本以及 7 个参考株进行了 tuf 基因 412bp 片段的基因分型和凝固酶检测。此外,还对 rpoB 基因进行了基因分型,并对 pta 基因进行了 PCR-RFLP,以明确鉴定中间葡萄球菌群(SIG)的成员。API Staph 32 正确鉴定了所有金黄色葡萄球菌(11/11)、SIG 种的 83%(10/12)和凝固酶阴性种的 66%(19/29)。RapID 和 Staph-Zym 分别正确鉴定了凝固酶阳性株的 61%(14/23)和 0%(0/23),以及凝固酶阴性株的 10%(3/29)和 3%(1/29)。

结论

市售的表型种鉴定试验不能正确鉴定来自伴侣动物的凝固酶阴性和凝固酶阳性葡萄球菌种。使用 tuf 基因序列进行基因分型优于表型鉴定,可用于鉴定动物来源的葡萄球菌种。然而,建议使用 pta 基因或 rpoB 测序的 PCR-RFLP 作为区分 SIG 分离株的确认方法。

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