Diagnostic performance of a novel loop-mediated isothermal amplification (LAMP) assay targeting the apicoplast genome for malaria diagnosis in a field setting in sub-Saharan Africa

Malaria Journal

Research

Jan Jacobs¹  Jean-Pierre Van geertruyden²  Davis Nwakanma³  Joseph Okebe⁴  Eniyou C. Oriero⁴  Umberto D’Alessandro⁵ 

[1] Institute of Tropical Medicine, Antwerp, Belgium;Department of Microbiology and Immunology, KU Leuven, Leuven, Belgium;International Health Unit, University of Antwerp, Antwerp, Belgium;Medical Research Council Unit, PO Box 273, Banjul, The Gambia;Medical Research Council Unit, PO Box 273, Banjul, The Gambia;Institute of Tropical Medicine, Antwerp, Belgium;International Health Unit, University of Antwerp, Antwerp, Belgium;Medical Research Council Unit, PO Box 273, Banjul, The Gambia;Institute of Tropical Medicine, Antwerp, Belgium;London School of Hygiene and Tropical Medicine, London, UK;

关键词: Malaria diagnosis;    LAMP;    RDT;    Microscopy;    PCR;    Field-based;    Resource-limited settings;   

DOI  :  10.1186/s12936-015-0926-6

 received in 2015-06-22, accepted in 2015-09-29,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundNew diagnostic tools to detect reliably and rapidly asymptomatic and low-density malaria infections are needed as their treatment could interrupt transmission. Isothermal amplification techniques are being explored for field diagnosis of malaria. In this study, a novel molecular tool (loop-mediated isothermal amplification—LAMP) targeting the apicoplast genome of Plasmodium falciparum was evaluated for the detection of asymptomatic malaria-infected individuals in a rural setting in The Gambia.MethodsA blood was collected from 341 subjects (median age 9 years, range 1–68 years) screened for malaria. On site, a rapid diagnostic test (RDT, SD Bioline Malaria Antigen P.f) was performed, thick blood films (TBF) slides for microscopy were prepared and dry blood spots (DBS) were collected on Whatman® 903 Specimen collection paper. The TBF and DBS were transported to the field laboratory where microscopy and LAMP testing were performed. The latter was done on DNA extracted from the DBS using a crude (methanol/heating) extraction method. A laboratory-based PCR amplification was done on all the samples using DNA extracted with the Qiagen kit and its results were taken as reference for all the other tests.ResultsPlasmodium falciparum malaria prevalence was 37 % (127/341) as detected by LAMP, 30 % (104/341) by microscopy and 37 % (126/341) by RDT. Compared to the reference PCR method, sensitivity was 92 % for LAMP, 78 % for microscopy, and 76 % for RDT; specificity was 97 % for LAMP, 99 % for microscopy, and 88 % for RDT. Area under the receiver operating characteristic (ROC) curve in comparison with the reference standard was 0.94 for LAMP, 0.88 for microscopy and 0.81 for RDT. Turn-around time for the entire LAMP assay was approximately 3 h and 30 min for an average of 27 ± 9.5 samples collected per day, compared to a minimum of 10 samples an hour per operator by RDT and over 8 h by microscopy.ConclusionThe LAMP assay could produce reliable results the same day of the screening. It could detect a higher proportion of low density malaria infections than the other methods tested and may be used for large campaigns of systematic screening and treatment.

【 授权许可】

CC BY   
© Oriero et al. 2015