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Title: Development of a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) for clinical detection of Plasmodium falciparum gametocytes
Authors: Sureemas Buates
Sirasate Bantuchai
Jetsumon Sattabongkot
Eun Taek Han
Takafumi Tsuboi
Rachanee Udomsangpetch
Jeeraphat Sirichaisinthop
Peerapan Tan-ariya
Mahidol University
Armed Forces Research Institute of Medical Sciences, Thailand
Kangwon National University, College of Medicine
Ehime University
Vector Borne Disease Training Center
Keywords: Immunology and Microbiology;Medicine
Issue Date: 1-Sep-2010
Citation: Parasitology International. Vol.59, No.3 (2010), 414-420
Abstract: Plasmodium falciparum gametocytes are usually present in peripheral blood at a very low level, thus requiring a sensitive assay detection method. In this study, reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was developed for clinical detection of P. falciparum gametocytes. Transcripts of Pfs16 of sexually committed ring and Pfs25 of mature gametocytes were detected by RT-LAMP in 82 clinical blood samples using nested RT-PCR as a gold standard. RT-LAMP demonstrated a detection limit of 1 parasitized red blood cell (RBC)/500μl of blood for both Pfs16 and Pfs25 transcripts. For Pfs16 transcript, RT-LAMP detected all 30 samples positive by nested RT-PCR (100% sensitivity) and 1 in 52 samples negative by nested RT-PCR (98.1% specificity). For Pfs25 transcript, RT-LAMP detected all 15 samples positive by nested RT-PCR (100% sensitivity) and none of 67 samples negative by nested RT-PCR (100% specificity). Negative predictive value (NPV) and positive predictive value (PPV) of RT-LAMP for detection of Pfs16 transcript were 100% and 96.8%, respectively, and 100% for both when employing Pfs25 transcript. Detection rate of Pfs16 and Pfs25 transcripts by RT-LAMP in microscopically gametocyte-negative samples was 91.7% and 29.2%, respectively. Compared with nested RT-PCR, RT-LAMP had a higher sensitivity but similar specificity, with the advantage of a shorter assay time. As RT-LAMP requires very basic instruments and the results can be obtained by visual inspection, this technique provides a simple and reliable tool for epidemiological studies of malaria transmission and in gametocyte-targeted control programmes. © 2010 Elsevier Ireland Ltd.
ISSN: 13835769
Appears in Collections:Scopus 2006-2010

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