Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts

Public Deposited
Analytics

Downloadable Content

Download PDF
Request Version for Screen Reader
Creator
  • Potlapalli, Varun R.
    • Affiliation: School of Medicine
  • Muller, Meredith S.
    • Affiliation: School of Medicine
  • Ngasala, Billy
    • Other Affiliation: Muhimbili University of Health and Allied Sciences
  • Ali, Innocent Mbulli
    • Other Affiliation: University of Dschang
  • Na, Yu Bin
    • Affiliation: School of Medicine
  • Williams, Danielle R.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Kharabora, Oksana
    • Affiliation: School of Medicine
  • Chhetri, Srijana
    • Affiliation: School of Medicine
  • Liu, Mei S.
    • Affiliation: School of Medicine
  • Carey-Ewend, Kelly
    • Affiliation: Gillings School of Global Public Health
  • Lin, Feng-Chang
    • Affiliation: Gillings School of Global Public Health
  • Mathias, Derrick
    • Other Affiliation: University of Florida
  • Tarimo, Brian B.
    • Other Affiliation: Ifakara Health Institute-Bagamoyo Office
  • Juliano, Jonathan J.
    • Affiliation: Gillings School of Global Public Health
  • Parr, Jonathan B.
    • Other Affiliation: School of Medicine
  • Lin, Jessica T.
    • Other Affiliation: School of Medicine
Abstract
  • Plasmodium ovale curtisi (Poc) and Plasmodium ovale wallikeri (Pow) represent distinct non-recombining Plasmodium species that are increasing in prevalence in sub-Saharan Africa. Though they circulate sympatrically, co-infection within human and mosquito hosts has rarely been described. Separate 18S rRNA real-time PCR assays that detect Poc and Pow were modified to allow species determination in parallel under identical cycling conditions. The lower limit of detection was 0.6 plasmid copies/μL (95% CI 0.4–1.6) for Poc and 4.5 plasmid copies/μL (95% CI 2.7–18) for Pow, or 0.1 and 0.8 parasites/μL, respectively, assuming 6 copies of 18s rRNA per genome. However, the assays showed cross-reactivity at concentrations greater than 103 plasmid copies/μL (roughly 200 parasites/μL). Mock mixtures were used to establish criteria for classifying mixed Poc/Pow infections that prevented false-positive detection while maintaining sensitive detection of the minority ovale species down to 100 copies/μL (<1 parasite/μL). When the modified real-time PCR assays were applied to field-collected blood samples from Tanzania and Cameroon, species identification by real-time PCR was concordant with nested PCR in 19 samples, but additionally detected two mixed Poc/Pow infections where nested PCR detected a single Po species. When real-time PCR was applied to oocyst-positive Anopheles midguts saved from mosquitoes fed on P. ovale-infected persons, mixed Poc/Pow infections were detected in 11/14 (79%). Based on these results, 8/9 P. ovale carriers transmitted both P. ovale species to mosquitoes, though both Po species could only be detected in the blood of two carriers. The described real-time PCR approach can be used to identify the natural occurrence of mixed Poc/Pow infections in human and mosquito hosts and reveals that such co-infections and co-transmission are likely more common than appreciated.
Date of publication
Keyword
DOI
Identifier
Resource type
  • Article
Rights statement
  • In Copyright
License
  • Attribution 4.0 International
Journal title
  • PLOS Neglected Tropical Diseases
Journal volume
  • 17
Journal issue
  • 12
Page start
  • e0011274
Language
  • English
Version
  • Publisher
ISSN
  • 1935-2735
Publisher
  • Public Library of Science

Relations

Parents:

This work has no parents.

In Collection:

Items

Thumbnail Title Date Uploaded Visibility Actions
file details journal.pntd.0011274.pdf 2024-01-03 Public Download