{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T00:28:30Z","timestamp":1783643310323,"version":"3.55.0"},"reference-count":45,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T00:00:00Z","timestamp":1741046400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Spanish Strategic Health Action (AESI-ISCIII)","award":["PI22CIII\/00033"],"award-info":[{"award-number":["PI22CIII\/00033"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background\/Objectives: Many tropical diseases such as malaria, Chagas, human African Trypanosomiasis, and Lymphatic filariasis coexist in endemic countries, affecting more than 1 billion people worldwide, and are recognised as major global vector-borne diseases. Tackling this disease requires an accurate diagnosis that is sensitive, specific, and rapid. This study aimed to describe and validate a new highly sensitive and specific multiple-analysis system that can effectively detect numerous etiological agents in a single test. Methods: A total of 230 human blood samples were assessed retrospectively for parasite characterisation, as well as 58 stool samples from non-human primates. Primers and probes were designed in the small subunit ribosomal RNA gene, except for Plasmodium spp., for which the novel target was Cytochrome Oxidase Subunit 1. Results: The analytical specificity of the presented method was 100%, with no unspecific amplifications or cross-reactions with other blood parasitic diseases. The detection limit obtained was between 0.6 and 3.01 parasites\/\u00b5L for Plasmodium species, 1.8 parasites\/mL for Trypanosomatidae, and 2 microfilariae\/mL in the case of Filariae. The sensitivity, specificity, predictive values, and kappa coefficient reached almost 100%, except for Filariae, whose sensitivity dropped to 93.9% and whose negative predicted value dropped to 89.5%. The operational features described a turnaround and a hands-on time shorter than the compared methods with a lower cost per essay. Conclusions: This work presents a cost-effective and highly sensitive multiplexed tool (RT-PCR-bp) capable of performing simultaneous detection for blood parasitic diseases using specific fluorescence probes, enabling the diagnosis of low parasite loads and coinfections.<\/jats:p>","DOI":"10.3390\/diagnostics15050620","type":"journal-article","created":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T13:31:31Z","timestamp":1741095091000},"page":"620","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development and Assessment of a Multiple-Analysis System for Diagnosing Malaria and Other Blood Parasite Infections in Humans and Non-Human Primates"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-7532-2774","authenticated-orcid":false,"given":"\u00c1ngela","family":"Ceballos-Caro","sequence":"first","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7484-6987","authenticated-orcid":false,"given":"V\u00edctor","family":"Ant\u00f3n-Berenguer","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"},{"name":"Department of Microbiology and Parasitology, Hospital Universitario Severo Ochoa, 28914 Legan\u00e9s, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Lanza","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justinn","family":"Renelies-Hamilton","sequence":"additional","affiliation":[{"name":"Section for Ecology and Evolution, Department of Biology, University of Copenhagen, 2200 Copenhagen, Denmark"},{"name":"Jane Goodall Institute Spain in Senegal, Dindefelo Biological Station, Dindefelo 26005, Senegal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8677-0668","authenticated-orcid":false,"given":"Amanda","family":"Barciela","sequence":"additional","affiliation":[{"name":"Jane Goodall Institute Spain in Senegal, Dindefelo Biological Station, Dindefelo 26005, Senegal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5963-8824","authenticated-orcid":false,"given":"Pamela C.","family":"K\u00f6ster","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"},{"name":"Women for Africa Foundation, 28046 Madrid, Spain"},{"name":"Faculty of Health Sciences, Alfonso X El Sabio University (UAX), 28091 Villanueva de la Ca\u00f1ada, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4015-8553","authenticated-orcid":false,"given":"David","family":"Carmena","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"},{"name":"CIBERINFEC, ISCIII\u2014CIBER Infectious Diseases, Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2597-3100","authenticated-orcid":false,"given":"Mar\u00eda","family":"Flores-Ch\u00e1vez","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emeline","family":"Chanove","sequence":"additional","affiliation":[{"name":"Clinique V\u00e9t\u00e9rinaire du Val d\u2019Arve, 74930 Reignier-Esery, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1903-6711","authenticated-orcid":false,"given":"Jos\u00e9 Miguel","family":"Rubio","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory, National Microbiology Centre, Instituto de Salud Carlos III, 28220 Majadahonda, Spain"},{"name":"CIBERINFEC, ISCIII\u2014CIBER Infectious Diseases, Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,4]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2023). 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