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

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...

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Published in:PLOS Neglected Tropical Diseases
Main Authors: Varun R Potlapalli, Meredith S Muller, Billy Ngasala, Innocent Mbulli Ali, Yu Bin Na, Danielle R Williams, Oksana Kharabora, Srijana Chhetri, Mei S Liu, Kelly Carey-Ewend, Feng-Chang Lin, Derrick Mathias, Brian B Tarimo, Jonathan J Juliano, Jonathan B Parr, Jessica T Lin
Format: Article in Journal/Newspaper
Language:English
Published: Public Library of Science (PLoS) 2023
Subjects:
Online Access:https://doi.org/10.1371/journal.pntd.0011274
https://doaj.org/article/21e27b4171524e07bfd3e53cd491ce12
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spelling ftdoajarticles:oai:doaj.org/article:21e27b4171524e07bfd3e53cd491ce12 2024-02-04T09:58:37+01:00 Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts. Varun R Potlapalli Meredith S Muller Billy Ngasala Innocent Mbulli Ali Yu Bin Na Danielle R Williams Oksana Kharabora Srijana Chhetri Mei S Liu Kelly Carey-Ewend Feng-Chang Lin Derrick Mathias Brian B Tarimo Jonathan J Juliano Jonathan B Parr Jessica T Lin 2023-12-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0011274 https://doaj.org/article/21e27b4171524e07bfd3e53cd491ce12 EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0011274 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0011274 https://doaj.org/article/21e27b4171524e07bfd3e53cd491ce12 PLoS Neglected Tropical Diseases, Vol 17, Iss 12, p e0011274 (2023) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2023 ftdoajarticles https://doi.org/10.1371/journal.pntd.0011274 2024-01-07T01:41:23Z 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. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 17 12 e0011274
institution Open Polar
collection Directory of Open Access Journals: DOAJ Articles
op_collection_id ftdoajarticles
language English
topic Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
spellingShingle Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
Varun R Potlapalli
Meredith S Muller
Billy Ngasala
Innocent Mbulli Ali
Yu Bin Na
Danielle R Williams
Oksana Kharabora
Srijana Chhetri
Mei S Liu
Kelly Carey-Ewend
Feng-Chang Lin
Derrick Mathias
Brian B Tarimo
Jonathan J Juliano
Jonathan B Parr
Jessica T Lin
Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts.
topic_facet Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
description 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.
format Article in Journal/Newspaper
author Varun R Potlapalli
Meredith S Muller
Billy Ngasala
Innocent Mbulli Ali
Yu Bin Na
Danielle R Williams
Oksana Kharabora
Srijana Chhetri
Mei S Liu
Kelly Carey-Ewend
Feng-Chang Lin
Derrick Mathias
Brian B Tarimo
Jonathan J Juliano
Jonathan B Parr
Jessica T Lin
author_facet Varun R Potlapalli
Meredith S Muller
Billy Ngasala
Innocent Mbulli Ali
Yu Bin Na
Danielle R Williams
Oksana Kharabora
Srijana Chhetri
Mei S Liu
Kelly Carey-Ewend
Feng-Chang Lin
Derrick Mathias
Brian B Tarimo
Jonathan J Juliano
Jonathan B Parr
Jessica T Lin
author_sort Varun R Potlapalli
title Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts.
title_short Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts.
title_full Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts.
title_fullStr Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts.
title_full_unstemmed Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts.
title_sort real-time pcr detection of mixed plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts.
publisher Public Library of Science (PLoS)
publishDate 2023
url https://doi.org/10.1371/journal.pntd.0011274
https://doaj.org/article/21e27b4171524e07bfd3e53cd491ce12
geographic Arctic
geographic_facet Arctic
genre Arctic
genre_facet Arctic
op_source PLoS Neglected Tropical Diseases, Vol 17, Iss 12, p e0011274 (2023)
op_relation https://doi.org/10.1371/journal.pntd.0011274
https://doaj.org/toc/1935-2727
https://doaj.org/toc/1935-2735
1935-2727
1935-2735
doi:10.1371/journal.pntd.0011274
https://doaj.org/article/21e27b4171524e07bfd3e53cd491ce12
op_doi https://doi.org/10.1371/journal.pntd.0011274
container_title PLOS Neglected Tropical Diseases
container_volume 17
container_issue 12
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