Abstract

BackgroundAs malaria transmission decreases, the proportion of infections that are asymptomatic at any given time increases. This poses a challenge for diagnosis as routinely used rapid diagnostic tests (RDTs) miss asymptomatic malaria cases with low parasite densities due to poor sensitivity. Yet, asymptomatic infections can contribute to onward transmission of malaria and therefore act as infectious reservoirs and perpetuate malaria transmission. This study compared the performance of RDTs to loop-mediated isothermal amplification (LAMP) in the diagnosis of malaria during reactive active case detection surveillance.MethodsAll reported malaria cases in the Engela Health District of Namibia were traced back to their place of residence and persons living within the four closest neighbouring houses to the index case (neighbourhood) were tested for malaria infection with RDTs and dried blood spots (DBS) were collected. LAMP and nested PCR (nPCR) were carried out on all RDTs and DBS. The same procedure was followed in randomly selected control neighbourhoods.ResultsSome 3151 individuals were tested by RDT, LAMP and nPCR. Sensitivity of RDTs and LAMP were 9.30 and 95.50%, respectively, and specificities were 99.27 and 99.92%, respectively, compared to nPCR. LAMP carried out on collected RDTs showed a sensitivity and specificity of 95.35 and 99.85% compared to nPCR carried out on DBS. There were 2 RDT samples that were negative by LAMP but the corresponding DBS samples were positive by PCR.ConclusionThe study showed that LAMP had the equivalent performance as nPCR for the identification of Plasmodium falciparum infection. Given its relative simplicity to implement over more complex and time-consuming methods, such as PCR, LAMP is particularly useful in elimination settings where high sensitivity and ease of operation are important.

Highlights

  • As malaria transmission decreases, the proportion of infections that are asymptomatic at any given time increases

  • loop-mediated isothermal amplification (LAMP) was run on all rapid diagnostic tests (RDTs) and dry blood spot (DBS) samples. nested polymerase chain reaction (PCR) (nPCR) was run on all LAMP and RDT positive samples and 10% of the LAMP negative samples using DBS as the source of DNA

  • LAMP was carried out on 2642 RDTs and 2640 DBS samples. nPCR was carried out on 56 DBS samples corresponding to all positive samples by RDT and/or LAMP and 10% of negative DBS (Table 3)

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Summary

Introduction

The proportion of infections that are asymptomatic at any given time increases. This poses a challenge for diagnosis as routinely used rapid diagnostic tests (RDTs) miss asymptomatic malaria cases with low parasite densities due to poor sensitivity. This study compared the performance of RDTs to loop-mediated isothermal amplification (LAMP) in the diagnosis of malaria during reactive active case detection surveillance. Asymptomatic infections are usually associated with sub-patent infections, below the density detectable (< 50 parasites/μL) by current RDTs [2] Detection of these infections relies on more sensitive methods, such as polymerase chain reaction (PCR), which have a detection limit of 1–2 parasites/μL when extracted from a dry blood spot (DBS) [4, 6, 7]. Both LAMP and PCR have a high sensitivity as a result of the amplification of the detection signal, DNA

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