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Traditional molecular detection methods cannot distinguish Escherichia coli O157:H7 in viable or dead state. In this study, the loop-mediated is other mal amplification (LAMP) method combined with propidium monoazide (PMA) treatment was developed to selectively detect the viable Escherichia coli O157:H7, but not dead cells. Four primers, including outer primers and inner primers, were specially designed for recognizing six distinct sequences on the species-specific rfbE gene of Escherichia coli O157:H7 genome. PMA penetrated selectively through the compromised cell membranes and intercalated into DNA, amplification of DNA from dead cells was inhibited completely by 3.0 μg/mL PMA, whereas the DNA derived from viable cells was amplified remarkably within 1 h by PMA-LAMP. This study offers a novel molecular detection method to distinguish between the viable and dead cells.

Specific Detection of Viable E.coli O157:H7 Cells by Coupling Propidium Monoazide with Loop-Mediated Isothermal Amplification