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Development of a new real-time quantitative PCR assay for the detection of Staphylococcus aureus genotype B in cow milk, targeting the new gene adlb.

Type of publication Peer-reviewed
Publikationsform Original article (peer-reviewed)
Author Sartori C, Boss R, Ivanovic I, Graber H U,
Project New Insights into Mechanisms of Human-Bovine Host Jump of Staphylococcus aureus Clonal Cluster CC8
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Original article (peer-reviewed)

Journal Journal of dairy science
Volume (Issue) 100(10)
Page(s) 7834 - 7845
Title of proceedings Journal of dairy science
DOI 10.3168/jds.2017-12820

Abstract

The specific and reliable diagnosis of mastitis pathogens is essential for successful sanitation programs. The aim of the present study was to develop and evaluate a new real-time quantitative PCR (qPCR) assay for the very sensitive and specific detection of Staphylococcus aureus genotype B in cow milk samples. This mastitis pathogen is contagious and particularly prevalent in Switzerland and other central European countries. The new test is based on a rapid preparation of bacteria, followed by DNA isolation and qPCR for a unique target gene coding for the adhesion-like bovine protein (adlb). The inclusivity of the new target gene was 97% and the exclusivity 98%, meaning that other genotypes and bacterial species could be excluded with high reliability. The limit of detection of the new assay was 235 staphylococcal cell equivalents/mL of culture. The new test shows high intra- and interassay repeatability. Results are available within 2 d after sampling, allowing farmers and veterinarians to apply sanitation measures immediately. Based on the results of a preliminary field study, the diagnostic sensitivity and specificity of the new qPCR assay are 99 and 100%, respectively. The new analytical procedure is straightforward and can be applied for routine diagnostics.
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