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Journal of Clinical Microbiology, August 2007, p. 2521-2528, Vol. 45, No. 8
0095-1137/07/$08.00+0     doi:10.1128/JCM.02117-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Detection of Four Plasmodium Species by Genus- and Species-Specific Loop-Mediated Isothermal Amplification for Clinical Diagnosis{triangledown}

Eun-Taek Han,1,2 Risa Watanabe,1 Jetsumon Sattabongkot,3 Benjawan Khuntirat,3 Jeeraphat Sirichaisinthop,4 Hideyuki Iriko,5 Ling Jin,5 Satoru Takeo,1 and Takafumi Tsuboi1,5*

Cell-Free Science and Technology Research Center,1 Venture Business Laboratory, Ehime University, Matsuyama, Ehime 790-8577, Japan,5 Department of Parasitology, Kangwon National University College of Medicine, Chunchon 200-701, Korea,2 Department of Entomology, Armed Forces Research Institute of Medical Sciences, Bangkok 10400, Thailand,3 Vector Borne Disease Training Center, Pra Budhabat, Saraburi 18120, Thailand4

Received 16 October 2006/ Returned for modification 6 December 2006/ Accepted 31 May 2007

Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method, was developed for the clinical detection of four species of human malaria parasites: Plasmodium falciparum, P. vivax, P. malariae, and P. ovale. We evaluated the sensitivity and specificity of LAMP in comparison with the results of microscopic examination and nested PCR. LAMP showed a detection limit (analytical sensitivity) of 10 copies of the target 18S rRNA genes for P. malariae and P. ovale and 100 copies for the genus Plasmodium, P. falciparum, and P. vivax. LAMP detected malaria parasites in 67 of 68 microscopically positive blood samples (sensitivity, 98.5%) and 3 of 53 microscopically negative samples (specificity, 94.3%), in good agreement with the results of nested PCR. The LAMP reactions yielded results within about 26 min, on average, for detection of the genus Plasmodium, 32 min for P. falciparum, 31 min for P. vivax, 35 min for P. malariae, and 36 min for P. ovale. Accordingly, in comparison to the results obtained by microscopy, LAMP had a similar sensitivity and a greater specificity and LAMP yielded results similar to those of nested PCR in a shorter turnaround time. Because it can be performed with a simple technology, i.e., with heat-treated blood as the template, reaction in a water bath, and inspection of the results by the naked eye because of the use of a fluorescent dye, LAMP may provide a simple and reliable test for routine screening for malaria parasites in both clinical laboratories and malaria clinics in areas where malaria is endemic.

* Corresponding author. Mailing address: Cell-Free Science and Technology Research Center, Ehime University, Matsuyama, Ehime 790-8577, Japan. Phone: 81-89-927-8277. Fax: 81-89-927-9941. E-mail: tsuboi{at}ccr.ehime-u.ac.jp

{triangledown} Published ahead of print on 13 June 2007.

Journal of Clinical Microbiology, August 2007, p. 2521-2528, Vol. 45, No. 8
0095-1137/07/$08.00+0     doi:10.1128/JCM.02117-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Peyrefitte, C. N., Boubis, L., Coudrier, D., Bouloy, M., Grandadam, M., Tolou, H. J., Plumet, S. (2008). Real-Time Reverse-Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of Rift Valley Fever Virus. J. Clin. Microbiol. 46: 3653-3659 [Abstract] [Full Text]  


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