Development of a loop-mediated isothermal amplification assay for rapid detection of Yersinia enterocolitica via targeting a conserved locus
Abstract
Background and Objectives: Loop-mediated isothermal amplification is a novel nucleic acid amplification assay providing as a simple diagnostic tool for rapid identification of microbial diseases in developing countries. In this study, a LAMP assay was established for Yersinia enterocolitica, a leading cause of acute enterocolitis in young children.
Materials and Methods: LAMP assay was established with four primers targeting a specific locus for the detection of Y. enterocolitica. The assay was conducted at 65°C in thermo block for 90min. The sensitivity of LAMP was evaluated in com- parison to conventional PCR using pTZ57R containing the target locus. Finally, specificity was assessed using DNA from common enteropathogenic bacteria.
Results: Results showed that the sensitivity of LAMP assay was 44-copy number, which was 10-fold higher than that of PCR. No cross-reactivity was observed when testing against other enteropathogenic pathogens.
Conclusion: This study showed that LAMP assay is an alternative molecular diagnostic tool for infections with Y. enteroco- litica. In addition, this method may be useful in diagnosis at field or in laboratories without PCR machine.
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| Issue | Vol 7 No 4 (2015) | |
| Section | Articles | |
| Keywords | ||
| Yersinia enterocolitica Loop-mediated isothermal amplification (LAMP) specific locus | ||
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