Rapid Identification of Vibrio cholerae Bacteria using LAMP and PCR methods in Water and Wastewater

Abstract

Vibrio cholerae serogroup 01 is known as the cause of epidemic cholera. Cholera is a pandemic or epidemic infectious disease that only occurs in humans and if it is not paid attention, it will lead to death in 6 hours or less a week. Recent epidemics of cholera around the world make the rapid and reliable detection of Vibrio cholerae necessary more than ever. Therefore, its early, accurate and on-time detection seems essential to prevent its epidemic and the devastating effects. There are several methods for detecting Vibrio cholerae, such as culturing, immunoassay, PCR and Real-time PCR. All these methods are time consuming and require large numbers of bacteria in the initial sample, expensive laboratory equipment and professionals in this field. In this study, PCR method was studied and compared with LAMP method to detect Vibrio cholerae. Bacterial DNA was extracted according to standard methods and detected using specific primers in PCR machine based on thermal cycle program. In contrast, the specific primer sets were amplified by LAMP method, at a single temperature in a thermal block and created products were identified through the turbidity examined visually and using gel electrophoresis. The results of both methods in molecular detection of Vibrio cholerae showed that LAMP method is faster, cheaper and more specific and Vibrio can be detected in a shorter time, using a very simple and inexpensive heat block and at a single temperature of 62 °C, as well as, observing the turbidity resulted from gene amplification process in new ways compared with electrophoresis method in examining PCR products, instead of expensive PCR machine and multi-temperature thermal cycle program. This method can be used for quick, accurate and affordable molecular detection of Vibrio cholerae with wide application in diagnostic laboratories, laboratory in underdeveloped regions and mobile laboratories.