Document Detail


Simultaneous detection of mastitis pathogens, Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae by multiplex real-time polymerase chain reaction.
MedLine Citation:
PMID:  16162525     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The objective of this study was to develop a multiplex real-time polymerase chain reaction (PCR) method for simultaneous detection of Staphylococcus aureus, Streptococcus agalactiae, and Streptococcus uberis directly from milk. A genetic marker specific for Staph. aureus was used for primers and dual-labeled probe design. The target for Strep. agalactiae primers and dual-labeled probe was selected from the cfb gene encoding the Christie-Atkins-Munch-Petersen factor. The plasminogen activator gene was the target for primers and dual-labeled probe design for Strep. uberis. Quarter milk samples (n = 192) were analyzed by the multiplex real-time PCR assay and conventional microbiological methods. An additional 57 quarter milk samples were analyzed in a separate real-time PCR assay for Strep. agalactiae only. Using an overnight enrichment step, the real-time PCR technique correctly identified 96.4% of all quarter milk samples; 91.7% of Staph. aureus, 98.2% of Strep. agalactiae, and 100% of Strep. uberis. Results of conventional microbiological methods were used to determine the sensitivity and specificity of the multiplex real-time PCR procedure. The sensitivity of the procedure to correctly identify Staph. aureus, Strep. agalactiae, and Strep. uberis directly from milk was 95.5%, and the specificity was 99.6%. Results of this study indicate that the multiplex real-time PCR procedure has the potential to be a valuable diagnostic technique for simultaneous identification of Staph. aureus, Strep. agalactiae, and Strep. uberis directly from quarter milk samples.
Authors:
B E Gillespie; S P Oliver
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of dairy science     Volume:  88     ISSN:  1525-3198     ISO Abbreviation:  J. Dairy Sci.     Publication Date:  2005 Oct 
Date Detail:
Created Date:  2005-09-15     Completed Date:  2006-03-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985126R     Medline TA:  J Dairy Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3510-8     Citation Subset:  IM    
Affiliation:
Food Safety Center of Excellence, Department of Animal Science, The University of Tennessee, Knoxville 37996, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
DNA, Bacterial / analysis
Female
Genetic Markers
Mastitis, Bovine / microbiology*
Milk / microbiology*
Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Staphylococcus aureus / genetics,  isolation & purification*
Streptococcus / genetics,  isolation & purification*
Streptococcus agalactiae / genetics,  isolation & purification*
Chemical
Reg. No./Substance:
0/DNA, Bacterial; 0/Genetic Markers

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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