Publication: Simple, rapid and sensitive detection of Orientia tsutsugamushi by loop-isothermal DNA amplification
Issued Date
2008-12-01
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ISSN
00359203
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2-s2.0-54949158969
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Mahidol University
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SCOPUS
Bibliographic Citation
Transactions of the Royal Society of Tropical Medicine and Hygiene. Vol.102, No.12 (2008), 1239-1246
Suggested Citation
Daniel H. Paris, Stuart D. Blacksell, Paul N. Newton, Nicholas P.J. Day Simple, rapid and sensitive detection of Orientia tsutsugamushi by loop-isothermal DNA amplification. Transactions of the Royal Society of Tropical Medicine and Hygiene. Vol.102, No.12 (2008), 1239-1246. doi:10.1016/j.trstmh.2008.04.040 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/19267
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Title
Simple, rapid and sensitive detection of Orientia tsutsugamushi by loop-isothermal DNA amplification
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Abstract
We present a loop-mediated isothermal PCR assay (LAMP) targeting the groEL gene, which encodes the 60 kDa heat shock protein of Orientia tsutsugamushi. Evaluation included testing of 63 samples of contemporary in vitro isolates, buffy coats and whole blood samples from patients with fever. Detection limits for LAMP were assessed by serial dilutions and quantitation by real-time PCR assay based on the same target gene: three copies/μl for linearized plasmids, 26 copies/μl for VERO cell culture isolates, 14 copies/μl for full blood samples and 41 copies/μl for clinical buffy coats. Based on a limited sample number, the LAMP assay is comparable in sensitivity with conventional nested PCR (56 kDa gene), with limits of detection well below the range of known admission bacterial loads of patients with scrub typhus. This inexpensive method requires no sophisticated equipment or sample preparation, and may prove useful as a diagnostic assay in financially poor settings; however, it requires further prospective validation in the field setting. © 2008 Royal Society of Tropical Medicine and Hygiene.