Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis.

Authors:
Eun-Taek Han
Eun-Taek Han
South Korea
Risa Watanabe
Risa Watanabe
Kinki Central Hospital
Japan
Jetsumon Sattabongkot
Jetsumon Sattabongkot
Mahidol University
Thailand
Benjawan Khuntirat
Benjawan Khuntirat
Armed Forces Research Institute of Medical Sciences
Jeeraphat Sirichaisinthop
Jeeraphat Sirichaisinthop
Vector Borne Disease Training Center
Hideyuki Iriko
Hideyuki Iriko
Ehime University Graduate School of Medicine
Japan
Ling Jin
Ling Jin
College of Veterinary Medicine
United States
Satoru Takeo
Satoru Takeo
Ehime University
Japan

J Clin Microbiol 2007 Aug 13;45(8):2521-8. Epub 2007 Jun 13.

Cell-Free Science and Technology Research Center, Ehime University, Matsuyama, Ehime 790-8577, Japan.

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.

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Source
http://dx.doi.org/10.1128/JCM.02117-06DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1951264PMC

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August 2007
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