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A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria.

ABSTRACT: Among vector-borne diseases malaria is the leading cause of morbidity in the world, with more than 200 million cases per year and a large number of deaths. The techniques traditionally used for the detection of Plasmodium in humans and Anopheles mosquitoes include microscopy, IRMA, ELISA, antibody or molecular assays, and anopheline dissection. However, these techniques are limited by their requirement of skilled personnel, low sensitivity or long processing times. A PCR-based high-resolution melting (PCR-HRM) analysis was developed for the detection and identification of P. falciparum, P. vivax and P. malariae that infect humans and Anopheles. In 41 human samples PCR-HRM detected 14 samples positive for P. vivax, 17 for P. falciparum, three for P. malariae, three mixed infections for P. vivax/P. malariae and four negative samples. Whereas benchmarking assays of microscopy and nested PCR had false positive detections. Additionally, PCR-HRM was able to detect natural infection with Plasmodium spp. in An. darlingi and An. mattogrossensis. The PCR-HRM presented is the first single assay developed for the detection and identification of P. vivax, P. falciparum and/or P. malariae in human and Anopheles. This method improves on currently available assays as it is easy-to-use, rapid, sensitive and specific with a low risk of contamination.

SUBMITTER: Murillo E

PROVIDER: S-EPMC6368607 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria.

Murillo Enderson E Muskus Carlos C Agudelo Luz A LA Vélez Iván D ID Ruiz-Lopez Freddy F

Scientific reports 20190208 1

Among vector-borne diseases malaria is the leading cause of morbidity in the world, with more than 200 million cases per year and a large number of deaths. The techniques traditionally used for the detection of Plasmodium in humans and Anopheles mosquitoes include microscopy, IRMA, ELISA, antibody or molecular assays, and anopheline dissection. However, these techniques are limited by their requirement of skilled personnel, low sensitivity or long processing times. A PCR-based high-resolution me ...[more]

PMID: 30737420

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Project description:BackgroundAntibody responses against Anopheles salivary proteins can indicate individual exposure to bites of malaria vectors. The extent to which these salivary proteins are species-specific is not entirely resolved. Thus, a better knowledge of the diversity among salivary protein repertoires from various malaria vector species is necessary to select relevant genus-, subgenus- and/or species-specific salivary antigens. Such antigens could be used for quantitative (mosquito density) and qualitative (mosquito species) immunological evaluation of malaria vectors/host contact. In this study, salivary gland protein repertoires (sialomes) from several Anopheles species were compared using in silico analysis and proteomics. The antigenic diversity of salivary gland proteins among different Anopheles species was also examined.ResultsIn silico analysis of secreted salivary gland protein sequences retrieved from an NCBInr database of six Anopheles species belonging to the Cellia subgenus (An. gambiae, An. arabiensis, An. stephensi and An. funestus) and Nyssorhynchus subgenus (An. albimanus and An. darlingi) displayed a higher degree of similarity compared to salivary proteins from closely related Anopheles species. Additionally, computational hierarchical clustering allowed identification of genus-, subgenus- and species-specific salivary proteins. Proteomic and immunoblot analyses performed on salivary gland extracts from four Anopheles species (An. gambiae, An. arabiensis, An. stephensi and An. albimanus) indicated that heterogeneity of the salivary proteome and antigenic proteins was lower among closely related anopheline species and increased with phylogenetic distance.ConclusionThis is the first report on the diversity of the salivary protein repertoire among species from the Anopheles genus at the protein level. This work demonstrates that a molecular diversity is exhibited among salivary proteins from closely related species despite their common pharmacological activities. The involvement of these proteins as antigenic candidates for genus-, subgenus- or species-specific immunological evaluation of individual exposure to Anopheles bites is discussed.

Dataset Information

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria.

Publications

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria.

Similar Datasets

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