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Nationwide Surveillance of Pfhrp2 Exon 2 Diversity in Plasmodium falciparum Circulating in Symptomatic Malaria Patients Living in Ghana

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  • 1 Department of Immunology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, Ghana;
  • | 2 Department of Microbiology and Immunology, School of Medical Sciences, College of Health and Allied Sciences, University of Cape Coast, Cape Coast, Ghana;
  • | 3 National Malaria Control Program, Ghana Health Services, Accra, Ghana;
  • | 4 Department of Epidemiology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, Ghana;
  • | 5 Department of Biomedical Science, School of Allied Health Sciences, College of Allied Health Sciences, University of Cape Coast, Cape Coast, Ghana;
  • | 6 Directorate of Research, Innovation and Consultancy, University of Cape Coast, Cape Coast, Ghana;
  • | 7 Department of Bioinformatics and Genomics, University of North Carolina, Charlotte, North Carolina;
  • | 8 Department of Biological Sciences, University of North Carolina, Charlotte, North Carolina;
  • | 9 School of Data Science, University of North Carolina, Charlotte, North Carolina
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ABSTRACT.

Reports of increasing false-negative HRP2-based rapid diagnostic test results across Africa require constant monitoring of factors associated with these false-negative outcomes, as failure of this diagnostic tool will have severe consequences on malaria treatment and control programs. This study characterized the extent of genetic diversity in the Plasmodium falciparum histidine-rich protein 2 (Pfhrp2) gene in P. falciparum isolates from symptomatic malaria patients across the regions of Ghana. Exon 2 of Pfhrp2 was amplified from gDNA using polymerase chain reaction. All Pfhrp2-negative samples were subjected to Pf18S rRNA and Pfmsp2 gene amplifications. The amplified Pfhrp2 exon 2 fragments from clonal samples were sent for commercial Sanger sequencing. The type and number of PfHRP2 repeats, classified based on repeat types previously reported, were estimated from the sequence data and compared among geographical regions. About 81% (2,333/2,890) of the original microscopy positive dried blood spot (DBS) samples were available and used in this study. The Pfhrp2 exon 2 amplification was successful in 98.5% (2,297/2,333) of the tested samples, with band size ranging from 400 bp to 1,050 bp. A total of 13 out of the 24 previously reported repeat types were identified among the samples, with three samples lacking both type 2 and type 7 repeat motifs. This study suggested that the genetic diversity of Pfhrp2 exon 2 identified in P. falciparum circulating in symptomatic malaria patients in Ghana is unlikely to influence the sensitivity and specificity of HRP2 RDT-based diagnosis.

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Author Notes

Address correspondence to Linda E. Amoah, Department of Immunology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, Ghana. E-mail: lamoah@ug.edu.gh

Financial support: This work was funded by the Global Fund through the Ghana National Malaria Control Program.

Disclosure: Approval of the study was obtained from the Institutional Review Board (IRB) of the Noguchi Memorial Institute for Medical Research (NMIMR).

Disclaimer: All data generated or analyzed during this study are included in the manuscript and its supplementary files.

Authors’ addresses: Dorcas G. Bredu, Dickson Donu, and Linda E. Amoah, Department of Immunology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, Ghana, E-mails: gyamahdorcas11@gmail.com, ddonu100@gmail.com, and lamoah@ug.edu.gh. George K. Ahadzi, Department of Microbiology and Immunology, School of Medical Sciences, College of Health and Allied Sciences, University of Cape Coast, Cape Coast, Ghana, E-mail: george.ahadzi@stu.ucc.edu.gh. Nana Y. Peprah, Alexander Asamoah, George A. Asumah, and Keziah L. Malm, National Malaria Control Program, Accra, Ghana, E-mails: naya_pep@yahoo.com, lexislea@yahoo.com, georgeaduasumah@yahoo.com, and keziah.malm@ghsmail.org. Benjamin Abuaku, Department of Epidemiology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, Ghana, E-mail: babuaku@noguchi.ug.edu.gh. Kwame K. Asare, Department of Biomedical Science, School of Allied Health Sciences, College of Allied Health Sciences, University of Cape Coast, Cape Coast, Ghana, E-mail: kwamsare@hotmail.com. Dorcas Obiri-Yeboah, Department of Microbiology and Immunology, School of Medical Sciences, College of Health and Allied Sciences, University of Cape Coast, Cape Coast, Ghana, and Directorate of Research, Innovation and Consultancy, University of Cape Coast, Cape Coast, Ghana, E-mail: dobiri-yeboah@ucc.edu.gh. Colby T. Ford, Department of Bioinformatics and Genomics, University of North Carolina, Charlotte, NC, and Department of Biological Sciences, University of North Carolina, Charlotte, NC, E-mail: cford38@uncc.edu. Eugenia Lo, Department of Biological Sciences, University of North Carolina, Charlotte, NC, and School of Data Science, University of North Carolina, Charlotte, NC, E-mail: eugenia.lo@uncc.edu.

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