The development of loop-mediated isothermal amplification targeting alpha-tubulin DNA for the rapid detection of Plasmodium vivax

Malaria Journal

Research

Sookwan Jeong¹  Byoung-Kuk Na²  Sylvatrie-Danne Dinzouna-Boutamba³  Dong-Il Chung³  So-Young Joo³  Yeonchul Hong³  Hye-Won Yang³  Youn-Kyoung Goo³  Hyun-Hee Kong⁴  Won-Ki Lee⁵  Noboru Inoue⁶ 

[1] Department of Medicine, ROK Army Headquarters, Choongnam, Republic of Korea;Department of Parasitology and Institute of Health Sciences, Gyeongsang National University School of Medicine, 660-751, Jinju, Republic of Korea;Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine, 700-422, Daegu, Republic of Korea;Department of Parasitology, Dong-A University, College of Medicine, 602–714, Busan, Republic of Korea;Department of Preventive Medicine, Kyungpook National University Medical Center, Kyungpook National University School of Medicine, Daegu, Republic of Korea;National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Obihiro, Japan;

关键词: Malaria;    Plasmodium vivax;    Immunochromatographic rapid diagnostic tests;    Polymerase chain reaction;    Loop-mediated isothermal amplification;   

DOI  :  10.1186/1475-2875-13-248

 received in 2014-03-07, accepted in 2014-06-26,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundMalaria that is caused by Plasmodium vivax is the most widely distributed human malaria. Its recent resurgence in many parts of the world, including the Republic of Korea (ROK), emphasizes the importance of improved access to the early and accurate detection of P. vivax to reduce disease burden. In this study, a rapid and efficient loop-mediated isothermal amplification (LAMP)-based method was developed and validated using blood samples from malaria-suspected patients.MethodA LAMP assay targeting the α-tubulin gene for the detection of P. vivax was developed with six primers that recognize different regions of the target gene. The diagnostic performance of the α-tubulin LAMP assay was compared to three other tests: microscopic examinations, rapid diagnostic tests (RDTs), and nested polymerase chain reactions (PCRs) using 177 whole blood specimens obtained from ROK military personnel from May to December 2011.ResultsThe α-tubulin LAMP assay was highly sensitive with a detection limit of 100 copies of P. vivax α-tubulin gene per reaction within 50 min. It specifically amplified the target gene only from P. vivax. Validation of the α-tubulin LAMP assay showed that the assay had the highest sensitivity (P < 0.001 versus microscopy; P = 0.0023 versus RDT) when nested PCR was used as the gold standard and better agreement (concordance: 94.9%, kappa value: 0.865) with nested PCR than RDT and microscopy. A Receiver Operation Characteristics analysis showed that the diagnostic accuracy of the α-tubulin LAMP assay for vivax malaria was higher (Area Under Curve = 0.908) than RDT and microscopy.ConclusionThis study showed that the P. vivax α-tubulin LAMP assay, which can be used to diagnose early infections of vivax malaria, is an alternative molecular diagnostic tool and a point-of-care test that may help to prevent transmission in endemic areas.

【 授权许可】

Unknown   
© Dinzouna-Boutamba et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.