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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Detection and enumeration of Staphylococcus aureus from bovine milk samples by real-time polymerase chain reaction

Full text
Author(s):
Botaro, B. G. [1] ; Cortinhas, C. S. [2] ; Marco, L. V. [2] ; Moreno, J. F. G. [2] ; Silva, L. F. P. [2] ; Benites, N. R. [1] ; Santos, M. V. [2]
Total Authors: 7
Affiliation:
[1] Univ Sao Paulo, Sch Vet Med & Anim Sci, Dept Prevent Vet Med & Anim Hlth, BR-13635900 Sao Paulo - Brazil
[2] Univ Sao Paulo, Sch Vet Med & Anim Sci, Dept Anim Sci, BR-13635900 Sao Paulo - Brazil
Total Affiliations: 2
Document type: Journal article
Source: JOURNAL OF DAIRY SCIENCE; v. 96, n. 11, p. 6955-6964, NOV 2013.
Web of Science Citations: 13
Abstract

The aim of this study was to develop and evaluate a real-time quantitative PCR (qPCR)-based method to detect and quantify Staphylococcus aureus in bronopol-preserved milk samples from subclinical intramammary infections (IMI). Serial dilutions of milk artificially inoculated with Staph. aureus ATCC 29213 were used to establish a standard curve (cfu/mL) of the qPCR assay targeting the Staph. aureus thermonuclease-encoding gene nuc according to the strain plate count. The analytical sensitivity, specificity, and repeatability of the qPCR assay were determined. A total of 60 milk samples, collected from mammary quarters without abnormal appearance and with positive isolation of Staph. aureus, were submitted to both the qPCR protocol and Staph. aureus plate counting and results from both methods were compared. Staphylococcus aureus from bronopol-preserved, subclinical IMI milk samples were not accurately enumerated by qPCR compared with plate counting of the nonpreserved, raw milk sample. The detection limit of the qPCR protocol of inoculated Staph. aureus ATCC 29213 in bronopol-preserved milk samples was 1.04 x 10(1) cfu/mL. The qPCR protocol can be a high-throughput and rapid diagnostic assay to accurately detect Staph. aureus IMI from bronopol-preserved milk samples compared with a traditional culturing method. However, the proposed qPCR protocol is not accurate for counting of Staph. aureus in bronopol-preserved milk samples from naturally infected mammary glands. (AU)

FAPESP's process: 08/05077-3 - Counting and identification of mastitis causing agents by the polymerase chain reaction
Grantee:Bruno Garcia Botaro
Support Opportunities: Scholarships in Brazil - Doctorate