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    Multiplex PCR (polymerase chain reaction) assay for detection of E. coli O157:H7, Salmonella sp., Vibrio cholerae and Vibrio parahaemolyticus in spiked shrimps (Penaeus monodon)

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    Date
    2013
    Publisher
    © 2013 Pakistan Journal of Biological Sciences
    Author
    Fakruddin, M.D.
    Sultana, Mahmuda
    Ahmed, Monzur Morshed
    Chowdhury, Abhijit
    Choudhury, Naiyyum
    Metadata
    Show full item record
    URI
    http://hdl.handle.net/10361/7110
    Citation
    Fakruddin, M. D., Sultana, M., Ahmed, M. M., Chowdhury, A., & Choudhury, N. (2013). Multiplex PCR (polymerase chain reaction) assay for detection of E. coli O157:H7, salmonella sp., vibrio cholerae and vibrio parahaemolyticus in spiked shrimps (penaeus monodon). Pakistan Journal of Biological Sciences, 16(6), 267-274. doi:10.3923/pibs.2013.267.274
    Abstract
    The coastal aquaculture mainly shrimps constitute major export sector in Bangladesh and is increasingly shaped by international trade conditions and by national responses to those stringent quality and safety standards. PCR based validated methods for detection of major bacterial pathogens in shrimp might be very useful tool for ensuring quality and safety standards of exportable shrimps. The objective of this study was to evaluate overall performance (sensitivity and specificity) of the multiplex PCR assay for detection of Vibrio cholerae, Vibrio parahaemolyticus, Salmonella sp. and Escherichia coli O157:H7 from spiked shrimp samples. The targeted genes were ompW for V. cholerae, tdh for V. parahaemolyticus, sefA for Salmonella spp. and hlyEHEC for E. coli O157:H7. The genomic DNA was extracted by using standard method and amplified accordingly. Sensitivity of the assay was tested by inoculating the shrimp homogenate with viable cells of laboratory references strains (target pathogens). The genes were amplified individually both from culture homogenate and spiked samples. Twenty different uniplex and multiplex PCR assay were performed; the results showed that the sensitivity and specificity of multiplex PCR are comparable to that of the results of uniplex PCR for the samples. DNA extracted from shrimp samples spiked with non-target pathogen (Bacillus cereus, Shigella flexneri and Staphylococcus aureus) yielded negative results.
    Keywords
    E. coli O157:H7; Multiplex PCR assay; Salmonella spp; Spiked shrimp; Vibrio cholerae; Vibrio parahaemolyticus
     
    Description
    This article was published in the Pakistan Journal of Biological Sciences [© 2013 Asian Network for Scientific Information] and the definite version is available at : http://dx.doi.org/10.3923/pjbs.2013.267.274. The Journal's website is at: http://scialert.net/abstract/index.php?doi=pjbs.2013.267.274
    Publisher Link
    http://scialert.net/abstract/index.php?doi=pjbs.2013.267.274
    DOI
    http://dx.doi.org/10.3923/pjbs.2013.267.274
    Department
    Department of Mathematics and Natural Sciences, BRAC University
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