Field Evaluation of a High Throughput Loop Mediated Isothermal Amplification Test for the Detection of Asymptomatic Plasmodium Infections in Zanzibar

Berit Aydin-Schmidt, Ulrika Morris, Xavier C Ding, Irina Jovel, Mwinyi I Msellem, Daniel Bergman, Atiqul Islam, Abdullah S Ali, Spencer Polley, Iveth J Gonzalez, Andreas Mårtensson, Anders Björkman, Berit Aydin-Schmidt, Ulrika Morris, Xavier C Ding, Irina Jovel, Mwinyi I Msellem, Daniel Bergman, Atiqul Islam, Abdullah S Ali, Spencer Polley, Iveth J Gonzalez, Andreas Mårtensson, Anders Björkman

Abstract

Background: New field applicable diagnostic tools are needed for highly sensitive detection of residual malaria infections in pre-elimination settings. Field performance of a high throughput DNA extraction system for loop mediated isothermal amplification (HTP-LAMP) was therefore evaluated for detecting malaria parasites among asymptomatic individuals in Zanzibar.

Methods: HTP-LAMP performance was evaluated against real-time PCR on 3008 paired blood samples collected on filter papers in a community-based survey in 2015.

Results: The PCR and HTP-LAMP determined malaria prevalences were 1.6% (95%CI 1.3-2.4) and 0.7% (95%CI 0.4-1.1), respectively. The sensitivity of HTP-LAMP compared to PCR was 40.8% (CI95% 27.0-55.8) and the specificity was 99.9% (CI95% 99.8-100). For the PCR positive samples, there was no statistically significant difference between the geometric mean parasite densities among the HTP-LAMP positive (2.5 p/μL, range 0.2-770) and HTP-LAMP negative (1.4 p/μL, range 0.1-7) samples (p = 0.088). Two lab technicians analysed up to 282 samples per day and the HTP-LAMP method was experienced as user friendly.

Conclusions: Although field applicable, this high throughput format of LAMP as used here was not sensitive enough to be recommended for detection of asymptomatic low-density infections in areas like Zanzibar, approaching malaria elimination.

Conflict of interest statement

Iveth J. Gonzalez and Xavier C. Ding are employees of FIND. All other authors declare that no competing interests exist. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1. Detection of LAMP result under…
Fig 1. Detection of LAMP result under UV light.
N- negative, P- positive, NC-negative control, PC- positive control.
Fig 2. Distribution of quantitative PCR determined…
Fig 2. Distribution of quantitative PCR determined parasite densities.
pan- Pan plasmodium, Geometric mean values are indicated by horizontal lines. Geometric mean values are for PCR + 1.8 p/μL (range 0.1–770), for PCR+/ pan HTP-LAMP + 2.5 p/μL (range 0.2–770), for PCR+/ pan-HTP-LAMP– 1.4 p/μL (range 0.1–7) and for Chelex pan-LAMP + 3.4 p/μL (range 0.1–770).
Fig 3. Flow chart of high-throughput (HTP)-LAMP…
Fig 3. Flow chart of high-throughput (HTP)-LAMP evaluation.
Results of Pan Plasmodium HTP-LAMP, confirmed PCR and Chelex LAMP (Ch-LAMP) are summarized. spp-species.

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