Ultrasensitive qPCR-Based Detection of Plasmodium falciparum in Pregnant Women Using Dried Blood or Whole Blood Pellet Samples Processed through Different DNA Extraction Methods

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  • 1 Kilimanjaro Clinical Research Institute, Moshi, Tanzania;
  • | 2 National Institute for Medical Research, Tanga Research Centre, Tanga, Tanzania;
  • | 3 Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark; Department of Infectious Diseases, Copenhagen University Hospital (Rigshospitalet), Copenhagen, Denmark;
  • | 4 Kilimanjaro Christian Medical University College, Moshi, Tanzania

Highly sensitive molecular techniques for the detection of low-level Plasmodium falciparum parasitemia are highly useful for various clinical and epidemiological studies. However, differences in how blood samples are preserved, the quantity of blood stored, as well as genomic DNA extraction methods used may compromise the potential usefulness of these methodologies. This study compared diagnostic sensitivity based on microscopy and malaria rapid diagnostic tests (mRDTs), with quantitative polymerase chain reaction (qPCR) P. falciparum positivity of dried blood spots (DBS) or whole blood pellets (WBP) from pregnant women using different DNA extraction protocols (Chelex-saponin or a commercial kit). Samples from 129 pregnant women were analyzed, of which 13 were P. falciparum positive by mRDT and 5 by microscopy. By using extraction kit on WBP and on DBS, qPCR positivity was 27 (20.9%) and 16 (12.4%), respectively, whereas Chelex extraction on DBS only resulted in 4 (3.1%) P. falciparum positive samples. Thus, extraction using commercial kits greatly improve the likelihood of detecting P. falciparum infections.

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Author Notes

Address correspondence to Queen Saidi, Kilimanjaro Clinical Research Institute, Moshi, Tanzania. E-mail: queensaidi@gmail.com

Financial support: The results of this publication (TRIAL-2015-1076-IMPROVE) were generated with financial support from the EDCTP2 programme and the MRC/DFID/Wellcome Trust Joint Global Health Trials (JGHT) scheme to the Liverpool School of Tropical Medicine.

Authors’ addresses: Queen Saidi, Kilimanjaro Clinical Research Institute, Moshi, Tanzania, and Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark, and Department of Infectious Diseases, Copenhagen University Hospital (Rigshospitalet), Copenhagen, Denmark, E-mail: queensaidi@gmail.com. Daniel Minja, National Institute for Medical Research, Tanga Research Centre, Tanga, Tanzania, E-mail: minjartd@gmail.com. Judith Njau, Kilimanjaro Clinical Research Institute, Moshi, Tanzania, E-mail: judiestev26@gmail.com. Helle Hansson and Michael Alifrangis, Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark, and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark, E-mails: hellehan@sund.ku.dk and micali@sund.ku.dk. Reginald Kavishe, Kilimanjaro Clinical Research Institute, Moshi, Tanzania, and Kilimanjaro Christian Medical University College, Moshi, Tanzania, E-mail: rekavishe@yahoo.com.

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