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Research articles

ScienceAsia 44(2018): 365-370 |doi: 10.2306/scienceasia1513-1874.2018.44.365


Rapid and sensitive enumeration of total coliforms and Escherichia coli in water and foods by most-probable-number loop-mediated isothermal amplification (MPN-LAMP) method


Chonnikan Tongphroma, Sutima Preeprema, Mitsuaki Nishibushib, Varaporn Vuddhakula, Pimonsri Mittraparp-arthorna,*

 
ABSTRACT:     Total coliforms and Escherichia coli are bacterial indicators used to assess the microbiological qualities of water and foods. In this study, we developed a most-probable-number loop-mediated isothermal amplification (most probable number (MPN-LAMP)) method for the enumeration of total coliforms and E. coli in drinking water and vegetables. The LAMP primers were designed based on lacZ (lacZ-LAMP) and uidA (uidA-LAMP) genes for the detection of total coliforms and E. coli, respectively. In pure culture, the lacZ-LAMP and the uidA-LAMP were able to detect all of the 37 coliforms and 30 E. coli strains, respectively. In the artificially contaminated drinking water and vegetables, the combination of MPN and LAMP techniques (MPN-LAMP) was able to detect E. coli at 1 colony-forming unit (CFU)/100 ml and 5 CFU/g, respectively. From analysis of 33 drinking water and 46 vegetable samples, the total coliform detection obtained by the MPN-LAMP method was in agreement with the MPN detection technique. However, MPN-LAMP was more sensitive than the MPN technique for E. coli detection. Our findings revealed that the MPN-LAMP assay was more rapid and highly sensitive than MPN method. Thus this method could be considered for detection and enumeration of total coliforms and E. coli in the food industry

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a Department of Microbiology, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90110 Thailand
b Division of Human-Nature Dynamics, Centre for Southeast Asian Studies, Kyoto University, Sakyo-ku, Kyoto 606-8501 Japan

* Corresponding author, E-mail: pimonsri.m@psu.ac.th

Received 6 Jul 2018, Accepted 1 Oct 2018