Detection of Plasmodium falciparum by Light Microscopy, Loop-Mediated Isothermal Amplification, and Polymerase Chain Reaction on Day 3 after Initiation of Artemether–Lumefantrine Treatment for Uncomplicated Malaria in Bagamoyo District, Tanzania: A Comparative Trial

Lwidiko E. Mhamilawa Department of Women’s and Children’s Health, International Maternal and Child Health (IMCH), Uppsala University, Uppsala, Sweden;
Department of Parasitology and Medical Entomology, Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania;

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Berit Aydin-Schmidt Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden;

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Bruno P. Mmbando Tanga Centre, National Institute for Medical Research, Tanga, Tanzania

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Billy Ngasala Department of Women’s and Children’s Health, International Maternal and Child Health (IMCH), Uppsala University, Uppsala, Sweden;
Department of Parasitology and Medical Entomology, Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania;

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Ulrika Morris Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden;

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Microscopy-determined Plasmodium falciparum positivity rates exceeding 10% on day 3 after initiation of artemisinin-based combination therapy (ACT) is an important indicator of artemisinin resistance. However, microscopy does not detect low-density parasitemia, contrary to molecular tools such as loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR). We compared microscopy, LAMP, and PCR for detection of P. falciparum on day 3 after ACT in 256 patients with uncomplicated malaria in Bagamoyo District, Tanzania. Day 3 positivity rates were 0%, 84.8%, and 84.4% for each method, respectively. The sensitivity and specificity of LAMP against PCR was 100% (95% CI, 96.1–100) and 77.4% (95% CI, 58.9–90.4) when quantitative PCR-determined parasite densities were ≥ two parasites/µL. Loop-mediated isothermal amplification had comparable diagnostic accuracy to PCR and could potentially represent a field-friendly tool for determining day 3 positivity rates. However, what day 3 P. falciparum positivity determined using molecular methods represents needs to be further elucidated.

Author Notes

Address correspondence to Lwidiko E. Mhamilawa, International Maternal and Child Health (IMCH), Uppsala University, MTC-huset, Dag Hammarskjölds väg 14B, 2 tr, Uppsala 752 37, Sweden.‬‬‬ E-mail: lwidiko.edward@kbh.uu.se‬

Financial support: The Swedish International Development Agency [Bilateral Sida grant: Bil-Tz 16/9875007059] and the Swedish Research Council [Grant number: 2016-0577] provided financial assistance for this work.

Authors’ addresses: Lwidiko E. Mhamilawa and Billy Ngasala, Department of Parasitology and Medical Entomology, Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania, E-mails: lwidiko.edward@kbh.uu.se and bngasala70@yahoo.co.uk. Berit Aydin-Schmidt and Ulrika Morris, Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, E- mails: berit.schmidt@ki.se and ulrika.morris@ki.se. Bruno P. Mmbando, Tanga Centre, National Institute for Medical Research, Tanga, Tanzania, E-mail: b.mmbando@yahoo.com.

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