alexa Loop-Mediated Isothermal Amplification Assay Targeting the 47-Kda Gene of Orientia tsutsugamushi: A Rapid and Sensitive Alternative to Real-Time PCR
ISSN: 2161-0703

Journal of Medical Microbiology & Diagnosis
Open Access

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Research Article

Loop-Mediated Isothermal Amplification Assay Targeting the 47-Kda Gene of Orientia tsutsugamushi: A Rapid and Sensitive Alternative to Real-Time PCR

Erin Huber1,2, Darder Ji3, Lee Howell1,2, Zhiwen Zhang1,2, Hua-Wei Chen1,2, Wei-Mei Ching1,2 and Chien-Chung Chao1,2,*

1Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD, USA

2Uniformed Services University of the Health Sciences, Bethesda, MD, USA

3Research and Diagnostic Center, Centers for Disease Control, Department of Health, 161 Kun-Yang Street, Taipei 11579, Taiwan

*Corresponding Author:
Chien-Chung Chao
Viral and Rickettsial Diseases Department
Infectious Diseases Directorate
Naval Medical Research Center
Silver Spring, MD, USA
E-mail: [email protected]

Received Date: May 07, 2012; Accepted Date: June 05, 2012; Published Date: June 12, 2012

Citation: Huber E, Ji D, Howell L, Zhang Z, Chen HW, et al. (2012) Loop-Mediated Isothermal Amplification Assay Targeting the 47-Kda Gene of Orientia tsutsugamushi: A Rapid and Sensitive Alternative to Real-Time PCR. J Med Microb Diagn 1:112. doi: 10.4172/2161-0703.1000112

Copyright: © 2012 Huber E, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

A sensitive, specific and rapid diagnostic test for the detection of Orientia tsutsugamushi, the causative agent of scrub typhus, is necessary to accurately and promptly diagnose patients and ensure that they receive proper treatment. A Loop-Mediated Isothermal Amplification (LAMP) assay targeting the 47-kDa gene of tsutsugamushi was developed. The LAMP assay was capable of detecting eleven different strains of Orientia at levels comparable to that of the quantitative PCR based method of detection. Ten patient specimens, confirmed to be positive for Orientia by two different PCR methods, were tested and nine out of ten were determined to be positive by LAMP. In terms of specificity, the assay was able to differentiate between Orientia and other phylogenetically similar bacteria as well as mouse and human host DNA. In addition to being sensitive and specific, the LAMP reaction was completed in 1 hour, demonstrating that it is a highly time-efficient method of diagnosing scrub typhus.

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