alexa TaqMan PCR for detection of Vibrio cholerae O1, O139, non-O1, and non-O139 in pure cultures, raw oysters, and synthetic seawater.
Agri and Aquaculture

Agri and Aquaculture

Journal of Aquaculture Research & Development

Author(s): Lyon WJ

Abstract Share this page

Abstract Vibrio cholerae is recognized as a leading human waterborne pathogen. Traditional diagnostic testing for Vibrio is not always reliable, because this bacterium can enter a viable but nonculturable state. Therefore, nucleic acid-based tests have emerged as a useful alternative to traditional enrichment testing. In this article, a TaqMan PCR assay is presented for quantitative detection of V. cholerae in pure cultures, oysters, and synthetic seawater. Primers and probe were designed from the nonclassical hemolysin (hlyA) sequence of V. cholerae strains. This probe was applied to DNA from 60 bacterial strains comprising 21 genera. The TaqMan PCR assay was positive for all of the strains of V. cholerae tested and negative for all other species of Vibrio tested. In addition, none of the other genera tested was amplified with the TaqMan primers and probe used in this study. The results of the TaqMan PCR with raw oysters and spiked with V. cholerae serotypes O1 and O139 were comparable to those of pure cultures. The sensitivity of the assay was in the range of 6 to 8 CFU g(-1) and 10 CFU ml(-1) in spiked raw oyster and synthetic seawater samples, respectively. The total assay could be completed in 3 h. Quantification of the Vibrio cells was linear over at least 6 log units. The TaqMan probe and primer set developed in this study can be used as a rapid screening tool for the presence of V. cholerae in oysters and seawater without prior isolation and characterization of the bacteria by traditional microbiological methods.
This article was published in Appl Environ Microbiol and referenced in Journal of Aquaculture Research & Development

Relevant Expert PPTs

Relevant Speaker PPTs

  • Ahmed Massoud
    Chemical and biological remediation of lindane residue in aqueous media
    PPT Version | PDF Version
  • Muhamed Fakhri Omer
    Cathodoluminescence petrography for provenance studies of the sandstones of Ora Formation (Devonian-Carboniferous), Iraqi Kurdistan Region, Northern Iraq
    PPT Version | PDF Version
  • Mustafa M Hariri
    Importance of methods’ selection in the geosciences studies and exploration
    PPT Version | PDF Version
  • Esther Barreiro
    Biological Mechanisms of Muscle Mass Loss and Cachexia in Patients with COPD
    PDF Version
  • Afsar Rahbar
    Studies of the importance of Cytomegalovirus infection in breast cancer
    PPT Version | PDF Version
  • Jim Polarine
    Case studies of human fl ora and spore contamination in clean rooms
    PPT Version | PDF Version
  • Gulay Yelken Demirel
    Challenges in parenteral formulation development studies and an evaluation from QbD point of view
    PPT Version | PDF Version
  • Jianfeng Hong
    Extractable and leachable studies of parenteral infusion and transfusion products
    PPT Version | PDF Version
  • Yosef Yarden
    Classically, the 3’untranslated region (3’UTR) is that region in eukaryotic protein-coding genes from the translation termination codon to the polyA signal. It is transcribed as an integral part of the mRNA encoded by the gene. However, there exists another kind of RNA, which consists of the 3’UTR alone, without all other elements in mRNA such as 5’UTR and coding region. The importance of independent 3’UTR RNA (referred as I3’UTR) was prompted by results of artificially introducing such RNA species into malignant mammalian cells. Since 1991, we found that the middle part of the 3’UTR of the human nuclear factor for interleukin-6 (NF-IL6) or C/EBP gene exerted tumor suppression effect in vivo. Our subsequent studies showed that transfection of C/EBP 3’UTR led to down-regulation of several genes favorable for malignancy and to up-regulation of some genes favorable for phenotypic reversion. Also, it was shown that the sequences near the termini of the C/EBP 3’UTR were important for its tumor suppression activity. Then, the C/EBP 3’UTR was found to directly inhibit the phosphorylation activity of protein kinase CPKC in SMMC-7721, a hepatocarcinoma cell line. Recently, an AU-rich region in the C/EBP 3’UTR was found also to be responsible for its tumor suppression. Recently we have also found evidence that the independent C/EBP 3’UTR RNA is actually exists in human tissues, such as fetal liver and heart, pregnant uterus, senescent fibroblasts etc. Through 1990’s to 2000’s, world scientists found several 3’UTR RNAs that functioned as artificial independent RNAs in cancer cells and resulted in tumor suppression. Interestingly, majority of genes for these RNAs have promoter-like structures in their 3’UTR regions, although the existence of their transcribed products as independent 3’UTR RNAs is still to be confirmed. Our studies indicate that the independent 3’UTR RNA is a novel non-coding RNA species whose function should be the regulation not of the expression of their original mRNA, but of some essential life activities of the cell as a whole.
    PPT Version | PDF Version
  • Arny L Blanchard
    Long-term environmental studies and stewardship in Alaska: A case study from Port Valdez
    PPT Version | PDF Version
  • Nwankwo Norbert Ikechukwu
    Direct computerized translation of biological data into biological information is now feasible: The gains of digital signal processing-based bioinformatics techniques
    PPT Version | PDF Version
  • Soha M Hamdy
    Biochemical studies on the effect of turmeric on breast cancer of rats
    PPT Version | PDF Version
  • George R. Bousfield
    Influence of Glycans on Biological Activities of Pituitary Gonadotropins: Follicle-Stimulating and Luteinizing Hormone
    PPT Version | PDF Version
  • Adel Nefzi
    “Adel Nefzi-Torrey-Pines-Institute-for-Molecular-Studies-USA-Diversity-Oriented-Synthesis-of-low-Molecular-Weight-Acyclic-and-Heterocyclic-Compounds-from-Resin-bound-Polyamides-Application-for-Drug-Discovery”
    PPT Version | PDF Version
  • Dalia Hussein Soliman
    “Dalia Hussein Soliman-Al-Azhar-University-Egypt-Design-synthesis-docking-and-QSAR-studies-of-novel-3-5-diaryl-pyrazole-derivatives-and-their-evaluation-as-antioxidants-and-as-Immunomodulators-inhibitors-of-TNF-α-IL-2-IL-6”
    PPT Version | PDF Version
Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords