Real-time PCR and NASBA for rapid and sensitive detection of Vibrio cholerae in ballast water

Enumeration of viable non-culturable Vibrio cholerae using propidium monoazide combined with quantitative PCR

“…This variable effect of PMA is a fatal weakness for the purpose of early detection. The environmental water samples usually contain few, if any, V. cholerae cells (Fykse et al, 2012). Because of the unculturable nature of VBNC cells, it is difficult to make a 10 8 /ml cell suspension for the PMA-qPCR procedure even after centrifugation.…”

Simple Visualized Detection Method of Virulence-Associated Genes of Vibrio cholerae by Loop-Mediated Isothermal Amplification

“…To date, many methods have been developed for effective detection of O1/O139 V. cholerae contamination in food. Compared with the conventional culture-based microbiological detection assays, molecular biology-based methods are more rapid and sensitive, such as PCR (Kumar et al, 2010;Zago et al, 2017), real-time PCR (Garrido-Maestu et al, 2015;Casasola-Rodriguez et al, 2018), multiplex PCR (Bwire et al, 2018;Vu et al, 2018), oligonucleotide array hybridization (Nasrabadi et al, 2017), strand displacement amplification (SDA) (Phillips et al, 2018), rolling circle amplification (RCA) (Osterberg et al, 2014), cross-priming amplification (CPA) (Zhang et al, 2015), and nucleic acid sequence-based amplification (NASBA) (Fykse et al, 2012). Nevertheless, these methods require expensive equipments, which limit their wide application, particularly in on-site testing and large-scale survey.…”

Detection and Occurrence of Indicator Organisms and Pathogens

“…Moreover, there was a positive correlation between 7 of the shellfish sites and water samples, with 3 sites being tested negative and 4 testing positive using both methods. cholerae in ballast waters (Fykse et al, 2012). Using both a 4 hr culturing step to enrich for viable cells, followed by qPCR was the most sensitive method, allow for detection of 1 CFU per 100 mL of ballast water, satisfying IMOguidelines.…”