• 1.

    Fried M, Muehlenbachs A, Duffy PE , 2012. Diagnosing malaria in pregnancy: an update. Expert Rev Anti Infect Ther 10: 11771187.

  • 2.

    Azizi SC, Chongwe G, Chipukuma H, Jacobs C, Zgambo J, Michelo C , 2018. Uptake of intermittent preventive treatment for malaria during pregnancy with sulphadoxine-pyrimethamine (IPTp-SP) among postpartum women in Zomba District, Malawi: a cross-sectional study. BMC Pregnancy Childbirth 18: 108.

    • Search Google Scholar
    • Export Citation
  • 3.

    Murphy SC, Shott JP, Parikh S, Etter P, Prescott WR, Stewart VA , 2013. Review article: malaria diagnostics in clinical trials. Am J Trop Med Hyg 89: 824839.

    • Search Google Scholar
    • Export Citation
  • 4.

    Tedla M , 2019. A focus on improving molecular diagnostic approaches to malaria control and elimination in low transmission settings: review. Parasite Epidemiol Control 6: e00107.

    • Search Google Scholar
    • Export Citation
  • 5.

    Zainabadi K, Nyunt MM, Plowe CV , 2019. An improved nucleic acid extraction method from dried blood spots for amplification of Plasmodium falciparum kelch13 for detection of artemisinin resistance. Malar J 18: 192.

    • Search Google Scholar
    • Export Citation
  • 6.

    Seesui K, Imtawil K, Chanetmahun P, Laummaunwai P, Boonmars T , 2018. An alternative method for extracting Plasmodium DNA from EDTA whole blood for malaria diagnosis. Korean J Parasitol 56: 2532.

    • Search Google Scholar
    • Export Citation
  • 7.

    Kobayashi T, Gamboa D, Ndiaye D, Cui L, Sutton PL, Vinetz JM , 2015. Malaria diagnosis across the International Centers of Excellence for Malaria Research: platforms, performance, and standardization. Am J Trop Med Hyg 93(Suppl):99.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 8.

    Roth JM, Korevaar DA, Leeflang MMG, Mens PF , 2016. Molecular malaria diagnostics: a systematic review and meta-analysis. Crit Rev Clin Lab Sci 53: 87105.

    • Search Google Scholar
    • Export Citation
  • 9.

    Choi E-H, Lee SK, Ihm C, Sohn Y-H , 2014. Rapid DNA extraction from dried blood spots on filter paper: potential applications in biobanking. Osong Public Health Res Perspect 5: 351357.

    • Search Google Scholar
    • Export Citation
  • 10.

    Miguel RB, Coura JR, Samudio F, Suarez-Mutis MC , 2013. Evaluation of three different DNA extraction methods from blood samples collected in dried filter paper in Plasmodium subpatent infections from the Amazon region in Brazil. Rev Inst Med Trop São Paulo 55: 205208.

    • Search Google Scholar
    • Export Citation
  • 11.

     Taylor BJ, Martin KA, Arango E, Agudelo OM, Maestre A, Yanow SK, 2011. Real-time PCR detection of Plasmodium directly from whole blood and filter paper samples. Malar J 1: 244.

  • 12.

    Dyan Hera Wiluyaningtias Y , 2015. Evaluation of five methods used to extract deoxyribonucleic acid (DNA) from human malaria parasitized blood spotted on the filter paper. Asian J Appl Sci 3: 429436.

    • Search Google Scholar
    • Export Citation
  • 13.

    Moyeh MN et al.2019. Comparison of the accuracy of four malaria diagnostic methods in a high transmission setting in coastal Cameroon. J Parasitol Res 2019. Available at: https://doi.org/10.1155/2019/1417967.

    • Search Google Scholar
    • Export Citation
  • 14.

    Simon N, Shallat J, Williams-Weitzikoski C, Harrington WE , 2020. Optimization of Chelex 100 resin-based extraction of genomic DNA from dried blood spots. Biol Methods Protoc 5: bpaa009.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15.

    Hofmann N, Mwingira F, Shekalaghe S, Robinson LJ, Mueller I, Felger I , 2015. Ultra-sensitive detection of Plasmodium falciparum by amplification of multi-copy subtelomeric targets. PLoS Med 12: 121.

    • Search Google Scholar
    • Export Citation
  • 16.

    Strøm GEA, Tellevik MG, Hanevik K, Langeland N, Blomberg B , 2014. Comparison of four methods for extracting DNA from dried blood on filter paper for PCR targeting the mitochondrial Plasmodium genome. Trans R Soc Trop Med Hyg 108: 488494.

    • Search Google Scholar
    • Export Citation
  • 17.

    Canier L et al.2015. Malaria PCR detection in Cambodian low-transmission settings: dried blood spots versus venous blood samples. Am J Trop Med Hyg 92: 573577.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 18.

    Singh UA, Kumari M, Iyengar S , 2018. Method for improving the quality of genomic DNA obtained from minute quantities of tissue and blood samples using Chelex 100 resin. Biol Proced Online 20. Available at: https://doi.org/10.1186/S12575-018-0077-6.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19.

    Minja DTR et al.2013. Plasmodium falciparum mutant haplotype infection during pregnancy associated with reduced birthweight, Tanzania. Emerg Infect Dis 19: 14461454.

    • Search Google Scholar
    • Export Citation
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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
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ABSTRACT.

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, P.O. Box 2236, 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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