Loop-mediated isothermal amplification (LAMP) assay for rapid detection of viable but non-culturable Vibrio cholerae O1

Author:

Chamanrokh Parastoo1,Colwell Rita R.1234,Huq Anwar12

Affiliation:

1. Maryland Pathogen Research Institute, University of Maryland, College Park, MD, USA.

2. Maryland Institute of Applied Environmental Health, University of Maryland, College Park, MD, USA.

3. Center of Bioinformatics and Computational Biology, University of Maryland Institute of Advanced Computer Studies, University of Maryland, College Park, MD, USA.

4. Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA.

Abstract

Vibrio cholerae, an important waterborne pathogen, is a rod-shaped bacterium that naturally exists in aquatic environments. When conditions are unfavorable for growth, the bacterium can undergo morphological and physiological changes to assume a coccoid morphology. This stage in its life cycle is referred to as viable but non-culturable (VBNC) because VBNC cells do not grow on conventional bacteriological culture media. The current study compared polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to detect and identify VBNC V. cholerae. Because it is difficult to detect and identify VBNC V. cholerae, the results of the current study are useful in showing that LAMP is more sensitive and rapid than PCR in detecting and identifying non-culturable, coccoid forms of V. cholerae. Furthermore, the LAMP method is effective in detecting and identifying very low numbers of coccoid VBNC V. cholerae in environmental water samples, with the added benefit of being inexpensive to perform.

Publisher

Canadian Science Publishing

Subject

Genetics,Molecular Biology,Applied Microbiology and Biotechnology,General Medicine,Immunology,Microbiology

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