Evaluating 18s-rRNA LAMP and selective whole genome amplification (sWGA) assay in detecting asymptomatic Plasmodium falciparum infections in blood donors

Malaria Journal

  ^{1    1    2    3    4} 

[1] 0000 0001 2322 8567, grid.413081.f, Department of Biomedical Sciences, School of Allied Health Sciences, University of Cape Coast, Cape Coast, Ghana;0000 0001 2322 8567, grid.413081.f, Department of Biomedical Sciences, School of Allied Health Sciences, University of Cape Coast, Cape Coast, Ghana;0000 0004 0606 5382, grid.10306.34, Wellcome Sanger Institute, CB10 1SA, Hinxton, UK;0000 0001 2322 8567, grid.413081.f, Department of Medical Laboratory Science, University of Cape Coast, Cape Coast, Ghana;0000 0004 1937 1485, grid.8652.9, School of Biological Sciences, University of Ghana, Legon, Ghana;

关键词: 18s-rRNA-LAMP;    Crude DNA LAMP;    Purified DNA LAMP;    Selective whole genome amplification;    P. falciparum;    Plasmodium;    Diagnostic indices;    Crude DNA extraction;   

DOI  :  10.1186/s12936-019-2850-7

来源: publisher

【 摘 要 】

BackgroundUndesirable consequences of donor Plasmodium falciparum parasitaemia on stored donor blood have been reported. Therefore, it is imperative that all prospective blood donors are screened for P. falciparum infections using sensitive techniques. In this study, the sensitivities of microscopy, rapid diagnostic test (RDT), loop-mediated isothermal amplification (LAMP) assay and selective whole genome amplification (sWGA) technique in detecting P. falciparum infections in blood donors was assessed.MethodsRandomly selected blood donors from 5 districts in Greater Accra Region of Ghana were screened for asymptomatic P. falciparum infections. Each donor sample was screened with SD Bioline RDT kit for P. falciparum histidine rich protein 2 and Plasmodium lactate dehydrogenase antigens, sWGA and 18s-rRNA LAMP. Crude DNA LAMP (crDNA-LAMP) was compared to purified DNA LAMP (pDNA-LAMP).ResultsA total of 771 blood donors were screened. The respective overall prevalence of P. falciparum in Ghana by microscopy, RDT, crDNA-LAMP, pDNA-LAMP and sWGA was 7.4%, 11.8%, 16.9%, 17.5% and 18.0%. Using sWGA as the reference test, the sensitivities of microscopy, RDT, crDNA-LAMP and pDNA-LAMP were 41.0% (95% CI 32.7–49.7), 65.5% (95% CI 56.9–73.3), 82.6% (95% CI 75.8–88.3) and 95.7% (95% CI 90.1–98.4), respectively. There was near perfect agreement between LAMP and sWGA (sWGA vs. crDNA-LAMP, κ = 0.87; sWGA vs. pDNA-LAMP, κ = 0.96), while crDNA-LAMP and pDNA-LAMP agreed perfectly (κ = 0.91). Goodness of fit test indicated non-significant difference between the performance of LAMP and sWGA (crDNA-LAMP vs. sWGA: x2 = 0.71, p = 0.399 and pDNA-LAMP vs. sWGA: x2 = 0.14, p = 0.707). Finally, compared to sWGA, the performance of LAMP did not differ in detecting sub-microscopic parasitaemia (sWGA vs. crDNA-LAMP: x2 = 1.12, p = 0.290 and sWGA vs. pDNA-LAMP: x2 = 0.22, p = 0.638).ConclusionsLAMP assay agreed near perfectly with sWGA with non-significant differences in their ability to detect asymptomatic P. falciparum parasitaemia in blood donors. Therefore, it is recommended that LAMP based assays are employed to detect P. falciparum infections in blood donors due to its high sensitivity, simplicity, cost-effectiveness and user-friendliness.

【 授权许可】

CC BY