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Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories
OBJECTIVE. To evaluate molecular tools to detect low-level parasitemia and the five species of Plasmodium that infect humans for use in control and elimination programs, and in reference laboratories. METHODS. We evaluated 145 blood samples from patients who tested positive by nested polymerase chai...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Organización Panamericana de la Salud
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8959250/ https://www.ncbi.nlm.nih.gov/pubmed/35355692 http://dx.doi.org/10.26633/RPSP.2022.11 |
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author | Aschar, Mariana Sanchez, Maria Carmen A. Costa-Nascimento, Maria de Jesus Farinas, Maria de Lourdes R. N. Hristov, Angélica D. Lima, Giselle F. M. C. Inoue, Juliana Levi, José E. Di Santi, Silvia M. |
author_facet | Aschar, Mariana Sanchez, Maria Carmen A. Costa-Nascimento, Maria de Jesus Farinas, Maria de Lourdes R. N. Hristov, Angélica D. Lima, Giselle F. M. C. Inoue, Juliana Levi, José E. Di Santi, Silvia M. |
author_sort | Aschar, Mariana |
collection | PubMed |
description | OBJECTIVE. To evaluate molecular tools to detect low-level parasitemia and the five species of Plasmodium that infect humans for use in control and elimination programs, and in reference laboratories. METHODS. We evaluated 145 blood samples from patients who tested positive by nested polymerase chain reaction (nPCR), from asymptomatic individuals and from the WHO Global Malaria Programme/United Kingdom National External Quality Assessment Service. Samples were assayed using the genus-specific RealStar(®) Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) and the RealStar(®) Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). The results from the molecular tests were compared with those from quantitative PCR (qPCR), nPCR and thick blood smear. RESULTS. The levels of parasitemia ranged from 1 to 518 000 parasites/µL, depending on the species. Compared with nPCR, alt-S&T had a sensitivity of 100%, except for identifying P. falciparum, for which the sensitivity was 93.94%. All samples positive by alt-Gen were also positive by nPCR. When comparing alt-Gen to qPCR, the sensitivity was 100% for P. vivax, P. malariae and P. falciparum. For all Plasmodium species, the correlation between cycle threshold values of alt-S&T and alt-Gen compared with qPCR was significant (P < 0.0001, Spearman’s test), with r = 0.8621 for alt-S&T and r = 0.9371 for alt-Gen. When all Plasmodium species were considered, there was a negative correlation between the level of parasitemia and real-time PCR cycle threshold values (P < 0.0001). In this study, only 2 of 28 samples from asymptomatic individuals were positive by thick blood smear; however, all 28 of these samples were positive by alt-S&T. CONCLUSIONS. The alt-Gen and alt-S&T assays are suitable for detecting submicroscopic infections for distinct epidemiological purposes, such as for use in surveys and reference laboratories, and screening in blood banks, which will contribute to global efforts to eliminate malaria. |
format | Online Article Text |
id | pubmed-8959250 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Organización Panamericana de la Salud |
record_format | MEDLINE/PubMed |
spelling | pubmed-89592502022-03-29 Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories Aschar, Mariana Sanchez, Maria Carmen A. Costa-Nascimento, Maria de Jesus Farinas, Maria de Lourdes R. N. Hristov, Angélica D. Lima, Giselle F. M. C. Inoue, Juliana Levi, José E. Di Santi, Silvia M. Rev Panam Salud Publica Original Research OBJECTIVE. To evaluate molecular tools to detect low-level parasitemia and the five species of Plasmodium that infect humans for use in control and elimination programs, and in reference laboratories. METHODS. We evaluated 145 blood samples from patients who tested positive by nested polymerase chain reaction (nPCR), from asymptomatic individuals and from the WHO Global Malaria Programme/United Kingdom National External Quality Assessment Service. Samples were assayed using the genus-specific RealStar(®) Malaria PCR Kit 1.0 (alt-Gen; altona Diagnostics) and the RealStar(®) Malaria Screen & Type PCR Kit (alt-S&T; altona Diagnostics). The results from the molecular tests were compared with those from quantitative PCR (qPCR), nPCR and thick blood smear. RESULTS. The levels of parasitemia ranged from 1 to 518 000 parasites/µL, depending on the species. Compared with nPCR, alt-S&T had a sensitivity of 100%, except for identifying P. falciparum, for which the sensitivity was 93.94%. All samples positive by alt-Gen were also positive by nPCR. When comparing alt-Gen to qPCR, the sensitivity was 100% for P. vivax, P. malariae and P. falciparum. For all Plasmodium species, the correlation between cycle threshold values of alt-S&T and alt-Gen compared with qPCR was significant (P < 0.0001, Spearman’s test), with r = 0.8621 for alt-S&T and r = 0.9371 for alt-Gen. When all Plasmodium species were considered, there was a negative correlation between the level of parasitemia and real-time PCR cycle threshold values (P < 0.0001). In this study, only 2 of 28 samples from asymptomatic individuals were positive by thick blood smear; however, all 28 of these samples were positive by alt-S&T. CONCLUSIONS. The alt-Gen and alt-S&T assays are suitable for detecting submicroscopic infections for distinct epidemiological purposes, such as for use in surveys and reference laboratories, and screening in blood banks, which will contribute to global efforts to eliminate malaria. Organización Panamericana de la Salud 2022-03-28 /pmc/articles/PMC8959250/ /pubmed/35355692 http://dx.doi.org/10.26633/RPSP.2022.11 Text en https://creativecommons.org/licenses/by-nc-nd/3.0/us/This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 IGO License, which permits use, distribution, and reproduction in any medium, provided the original work is properly cited. No modifications or commercial use of this article are permitted. In any reproduction of this article there should not be any suggestion that PAHO or this article endorse any specific organization or products. The use of the PAHO logo is not permitted. This notice should be preserved along with the article’s original URL. Open access logo and text by PLoS, under the Creative Commons Attribution-Share Alike 3.0 Unported license. |
spellingShingle | Original Research Aschar, Mariana Sanchez, Maria Carmen A. Costa-Nascimento, Maria de Jesus Farinas, Maria de Lourdes R. N. Hristov, Angélica D. Lima, Giselle F. M. C. Inoue, Juliana Levi, José E. Di Santi, Silvia M. Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories |
title | Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories |
title_full | Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories |
title_fullStr | Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories |
title_full_unstemmed | Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories |
title_short | Ultrasensitive molecular tests for Plasmodium detection: applicability in control and elimination programs and reference laboratories |
title_sort | ultrasensitive molecular tests for plasmodium detection: applicability in control and elimination programs and reference laboratories |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8959250/ https://www.ncbi.nlm.nih.gov/pubmed/35355692 http://dx.doi.org/10.26633/RPSP.2022.11 |
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