Multiplex PCR assay for species identification of bovine mastitis pathogens

• Published in: Journal of applied microbiology 2011-12, Vol.111 (6), p.1349-1356
• Main Authors: Shome, B.R, Das Mitra, S, Bhuvana, M, Krithiga, N, Velu, D, Shome, R, Isloor, S, Barbuddhe, S.B, Rahman, H
• Format: Article
• Language: English
• Subjects: Animals; bacteria; Bacteria - classification; Bacteria - genetics; Bacteria - isolation & purification; Biological and medical sciences; bovine mastitis; bulk milk; Cattle; detection; detection limit; DNA; DNA, Bacterial - analysis; Escherichia coli; Female; Food Contamination - analysis; Fundamental and applied biological sciences. Psychology; Limit of Detection; Mastitis, Bovine - diagnosis; Mastitis, Bovine - microbiology; Microbiology; milk; Milk - microbiology; multiplex PCR; Multiplex Polymerase Chain Reaction - methods; pathogen identification; pathogens; polymerase chain reaction; Sensitivity and Specificity; species identification; Species Specificity; Staphylococcus aureus; Staphylococcus chromogenes; Staphylococcus epidermidis; Staphylococcus haemolyticus; Staphylococcus sciuri; Staphylococcus simulans; Streptococcus agalactiae; Streptococcus dysgalactiae; Streptococcus uberis
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.2011.05169.x

Aim: To develop and evaluate a multiplex PCR (mPCR) assay for simultaneous detection of 10 bacterial species causing bovine mastitis namely, Staphylococcus aureus, Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus sciuri, Staphylococcus haemolyticus, Staphylococcus simulans, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis and Escherichia coli in milk. Methods and Results: A two‐tube mPCR assay was developed. The accuracy of the mPCR was evaluated using 56 standard reference strains and 705 strains comprising of E. coli (n = 99), staphylococci (n = 522) and streptococci (n = 84). The threshold of detection of the mPCR assay was 10 fg of genomic DNA and