Development of a loop-mediated isothermal amplification assay for detecting .

CONTEXT

Loop-mediated isothermal amplification (LAMP) may be used in the future to detect infecting microorganisms. LAMP assays exist for the endodontic pathogens and , but not yet for

AIM

To develop a LAMP assay for detecting .

SETTINGS AND DESIGN

It was an benchtop study.

SUBJECTS AND METHODS

The National Center for Biotechnology Information GenBank Basic Local Alignment Search Tool was used to identify a segment of the dipeptidyl peptidase 11 (DPP11) gene unique to . A primer design tool was used to generate six primers required for developing the LAMP assay. WarmStart Colorimetric LAMP 2X Master Mix was used to evaluate the LAMP assay, using purified DNA as a control.

STATISTICAL ANALYSIS USED

Statistical parameters for sensitivity and specificity.

RESULTS

The assay was performed in triplicate on pure DNA from and and on the DNA that was extracted from , , , and cells and diluted two-fold from 1/2 to 1/256. Assays for the diluted samples were performed in triplicate, and the contingency tables indicated the LAMP assay to be 82% sensitive and 90% specific for .

CONCLUSIONS

LAMP assay could be a highly sensitive and specific chairside detection method for .