{"doi":"10.17615/c7x5-2f78","title":"Real-time PCR detection of mixed Plasmodium ovale curtisi and wallikeri infections in human and mosquito hosts","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.","journal":"UNC Libraries","year":2024,"id":498814,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.96,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1081719,"name":"Varun R. Potlapalli","orcid":"0009-0001-0069-1353","position":1,"is_corresponding":false},{"id":866431,"name":"Brian B. Tarimo","orcid":"0000-0002-8097-8310","position":2,"is_corresponding":false},{"id":432697,"name":"Derrick Mathias","orcid":"0000-0002-8743-7285","position":3,"is_corresponding":false},{"id":787333,"name":"D. R. Williams","orcid":null,"position":4,"is_corresponding":false},{"id":647357,"name":"Meredith Muller","orcid":null,"position":5,"is_corresponding":false},{"id":1081720,"name":"Yu Bin Na","orcid":"0009-0001-4774-3480","position":6,"is_corresponding":false},{"id":527805,"name":"Oksana Kharabora","orcid":null,"position":7,"is_corresponding":false},{"id":647359,"name":"Srijana Chhetri","orcid":null,"position":8,"is_corresponding":false},{"id":270623,"name":"Jonathan B. Parr","orcid":"0000-0002-4465-9830","position":9,"is_corresponding":false},{"id":1082157,"name":"Mei S. Liu","orcid":null,"position":10,"is_corresponding":false},{"id":646275,"name":"Kelly Carey-Ewend","orcid":"0000-0002-3200-1644","position":11,"is_corresponding":false},{"id":1061560,"name":"Innocent Mbulli Ali","orcid":"0000-0002-1112-6376","position":12,"is_corresponding":false},{"id":381755,"name":"Feng‐Chang Lin","orcid":"0000-0002-2638-1775","position":13,"is_corresponding":false},{"id":270632,"name":"Jonathan J. Juliano","orcid":"0000-0002-0591-0850","position":14,"is_corresponding":false},{"id":300878,"name":"Billy Ngasala","orcid":"0000-0001-9300-7719","position":15,"is_corresponding":false},{"id":428587,"name":"Jessica T. Lin","orcid":"0000-0002-4516-723X","position":0,"is_corresponding":true}],"reference_count":0,"raw_metadata":null,"created_at":"2026-07-19T02:09:42.181447Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}