Evaluation and Improvement of LAMP Assays for Detection of Serogroups O26, O45, O103, O111, O121, O145, and O157

Overview

Journal Afr Health Sci

Date 2018 Jun 26

PMID 29937871

Citations 3

Authors

Deguo Wang

Affiliations

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Abstract

Objective: To evaluate existing LAMP assays for detection of the seven STEC serogroups, if necessary, to improve these assays and to promote their practical application.

Methods: Pure DNA extract from 23 strains reserved in our lab was used to evaluate the existing LAMP assays. The existing LAMP assays were modified via adding 1% tetramethylene sulfoxide and 5% dimethyl sulfoxide as well as optimizing reaction temperature.

Results: The detection limit of the modified LAMP assays was 0.1-1 pg per reaction, equivalent to 25-250 cfu per reaction, the non-specific amplification can completely be eliminated with optimal amplifying temperature, and the modified LAMP assays can specifically and sensitivly amplify targeted O serogroup-specific gene (wzx or wzy) of any concerned serogroup as commercial kit Isothermal Master Mix did.

Conclusion: In conclusion, the LAMP assays were highly susceptible to non-specific amplification caused by primer dimers, and these modified methods were free of non-specific amplification and can rapidly and reliably detect the seven major Shiga toxin-producing serogroups.

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