Validation and verification of LAMP, ISO, and VIDAS UP methods for detection of Escherichia coli O157:H7 in different food matrices.

Abstract

Escherichia coli O157:H7 is one of the most important food-borne pathogens to threaten public health. Cultural methods are used as a gold standard while they are laborious and time-consuming. Loop-mediated isothermal amplification (LAMP) method is an alternative method that became widely used for the detection of food-borne pathogens. The aim of this study was to evaluate the specificity and sensitivity of LAMP method for detection of E. coli O157:H7, also to compare detection performances with VIDAS UP and ISO (International Organization for Standardization) methods in different food matrices (beef meat, minced lamb meat, milk, cheese, apple puree, and soybean sprouts). E. coli O157:H7 were spiked in three different levels (high 4.58; medium 2.32; low 0.30 log(10) CFU/g-ml) to food matrices. Although there were no significant differences in terms of the specificity and sensitivity values among the three methods (p ? .05), it was determined that the highest specificity and sensitivity values obtained from the LAMP method. Sensitivity and specificity values of LAMP method were found as 0.997 and 0.988, for the ISO method were 0.989 and 0.971, and for the VIDAS UP method were 0.980 and 0.963, respectively. In milk samples, sensitivity and specificity values of the VIDAS UP method were significantly lower than LAMP and ISO methods (p < .05). However, there were no significant differences found for the other food matrices among the three methods (p > .05). It can be summarized from this study that specificity and sensitivity values of the LAMP method are equal or higher and less time-consuming than ISO and VIDAS UP methods. In conclusion, using a simple, fast, and inexpensive detection method, such as LAMP, especially in endemic regions or in an outbreak to control spreading of pathogens, is very important for public health.