4.7 Article

Short communication: Identification of coagulase-negative staphylococcus species from goat milk with the API Staph identification test and with transfer RNA-intergenic spacer PCR combined with capillary electrophoresis

Journal

JOURNAL OF DAIRY SCIENCE
Volume 95, Issue 12, Pages 7200-7205

Publisher

ELSEVIER SCIENCE INC
DOI: 10.3168/jds.2012-5747

Keywords

coagulase-negative Staphylococcus species; goat; transfer DNA-PCR; API 20 Staph identification test

Funding

  1. National Sheep Industry Improvement Center (Rockland, ME)
  2. American Dairy Goat Association (Spindale, NC)
  3. Dutch Dairy Board (Zoetermeer, the Netherlands)
  4. Dutch Association for Quality Assurance in Goat Farming (VKGN, Giessenburg, the Netherlands)

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Coagulase-negative staphylococci (CNS) are the most commonly isolated bacteria from goat milk, but they have often been identified with phenotypic methods, which may have resulted in misclassification. The aims of this paper were to assess the amount of misclassification of a phenotypic test for identifying CNS species from goat milk compared with transfer RNA intergenic spacer PCR (tDNA-PCR) followed by capillary electrophoresis, and to apply the tDNA-PCR technique on different capillary electrophoresis equipment. Milk samples were collected from 416 does in 5 Californian dairy goat herds on 3 occasions during lactation. In total, 219 CNS isolates were identified at the species level with tDNA-PCR and subjected to the API 20 Staph identification test kit (API Staph; bioMerieux, Durham, NC). If the same species was isolated multiple times from the same udder gland, only the first isolate was used for further analyses, resulting in 115 unique CNS isolates. According to the tDNA-PCR test, the most prevalent CNS species were Staphylococcus epidermidis, Staphylococcus caprae, and Staphylococcus simulans. Typeability with API staph was low (72%). Although the API Staph test was capable of identifying the majority of Staph. epidermidis and Staph. caprae isolates, sensitivity for identification of Staph. simulans was low. The true positive fraction was high for the 3 most prevalent species. It was concluded that the overall performance of API Staph in differentiating CNS species from goat milk was moderate to low, mainly because of the low typeability, and that genotypic methods such as tDNA-PCR are preferred.

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