Journal
MOLECULAR AND CELLULAR PROBES
Volume 33, Issue -, Pages 57-64Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.mcp.2017.03.004
Keywords
Bovine mastitis; Multiplex PCR assay; Latent class analysis
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Funding
- Higher Education Commission of Pakistan under the International Research Support Initiative Program (IRSIP) [266.22]
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For rapid and simultaneous detection of nine bovine mastitic pathogens, a sensitive and specific multiplex PCR assay was developed. The assay was standardized using reference strains and validated on mastitic milk cultures which were identified to species level based on 16S rRNA sequencing. Multiplex PCR assay also efficiently detected the target bacterial strains directly from milk. The detection limit of the assay was up to 50 pg for DNA isolated from pure cultures and 10(4) CFU/ml for spiked milk samples. As estimated by latent class analysis, the assay was sensitive up to 88% and specific up to 98% for targeted mastitic pathogens, compared with the bacterial culture method and the 16S rRNA sequence analysis. This novel molecular assay could be useful for monitoring and maintaining the bovine udder health, ensuring the bacteriological safety of milk, and conducting epidemiological studies. (C) 2017 Elsevier Ltd. All rights reserved.
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