KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum
Abstract Background The emergence and spread of anti-malarial resistance continues to hinder malaria control. Plasmodium falciparum, the species that causes most human malaria cases and most deaths, has shown resistance to almost all known anti-malarials. This anti-malarial resistance arises from th...
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ftdoajarticles:oai:doaj.org/article:dea477b7b63349e883bcd864bece77ea 2023-05-15T15:17:36+02:00 KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum Ana Alvarez-Fernandez María J. Bernal Isabel Fradejas Alexandra Martin Ramírez Noor Azian Md Yusuf Marta Lanza Shamilah Hisam Ana Pérez de Ayala José M. Rubio 2021-01-01T00:00:00Z https://doi.org/10.1186/s12936-020-03544-7 https://doaj.org/article/dea477b7b63349e883bcd864bece77ea EN eng BMC https://doi.org/10.1186/s12936-020-03544-7 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-020-03544-7 1475-2875 https://doaj.org/article/dea477b7b63349e883bcd864bece77ea Malaria Journal, Vol 20, Iss 1, Pp 1-8 (2021) Plasmodium falciparum KASP Resistance SNPs K13 MDR Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2021 ftdoajarticles https://doi.org/10.1186/s12936-020-03544-7 2022-12-31T07:50:15Z Abstract Background The emergence and spread of anti-malarial resistance continues to hinder malaria control. Plasmodium falciparum, the species that causes most human malaria cases and most deaths, has shown resistance to almost all known anti-malarials. This anti-malarial resistance arises from the development and subsequent expansion of Single Nucleotide Polymorphisms (SNPs) in specific parasite genes. A quick and cheap tool for the detection of drug resistance can be crucial and very useful for use in hospitals and in malaria control programmes. It has been demonstrated in different contexts that genotyping by Kompetitive Allele Specific PCR (KASP), is a simple, fast and economical method that allows a high-precision biallelic characterization of SNPs, hence its possible utility in the study of resistance in P. falciparum. Methods Three SNPs involved in most cases of resistance to the most widespread anti-malarial treatments have been analysed by PCR plus sequencing and by KASP (C580Y of the Kelch13 gene, Y86N of the Pfmdr1 gene and M133I of the Pfcytb gene). A total of 113 P. falciparum positive samples and 24 negative samples, previously analysed by PCR and sequencing, were selected for this assay. Likewise, the samples were genotyped for the MSP-1 and MSP-2 genes, and the Multiplicity of Infection (MOI) and parasitaemia were measured to observe their possible influence on the KASP method. Results The KASP results showed the same expected mutations and wild type genotypes as the reference method, with few exceptions that correlated with very low parasitaemia samples. In addition, two cases of heterozygotes that had not been detected by sequencing were found. No correlation was found between the MOI or parasitaemia and the KASP values of the sample. The reproducibility of the technique shows no oscillations between repetitions in any of the three SNPs analysed. Conclusions The KASP assays developed in this study were efficient and versatile for the determination of the Plasmodium genotypes related to ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 20 1 |
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English |
topic |
Plasmodium falciparum KASP Resistance SNPs K13 MDR Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Plasmodium falciparum KASP Resistance SNPs K13 MDR Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Ana Alvarez-Fernandez María J. Bernal Isabel Fradejas Alexandra Martin Ramírez Noor Azian Md Yusuf Marta Lanza Shamilah Hisam Ana Pérez de Ayala José M. Rubio KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum |
topic_facet |
Plasmodium falciparum KASP Resistance SNPs K13 MDR Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background The emergence and spread of anti-malarial resistance continues to hinder malaria control. Plasmodium falciparum, the species that causes most human malaria cases and most deaths, has shown resistance to almost all known anti-malarials. This anti-malarial resistance arises from the development and subsequent expansion of Single Nucleotide Polymorphisms (SNPs) in specific parasite genes. A quick and cheap tool for the detection of drug resistance can be crucial and very useful for use in hospitals and in malaria control programmes. It has been demonstrated in different contexts that genotyping by Kompetitive Allele Specific PCR (KASP), is a simple, fast and economical method that allows a high-precision biallelic characterization of SNPs, hence its possible utility in the study of resistance in P. falciparum. Methods Three SNPs involved in most cases of resistance to the most widespread anti-malarial treatments have been analysed by PCR plus sequencing and by KASP (C580Y of the Kelch13 gene, Y86N of the Pfmdr1 gene and M133I of the Pfcytb gene). A total of 113 P. falciparum positive samples and 24 negative samples, previously analysed by PCR and sequencing, were selected for this assay. Likewise, the samples were genotyped for the MSP-1 and MSP-2 genes, and the Multiplicity of Infection (MOI) and parasitaemia were measured to observe their possible influence on the KASP method. Results The KASP results showed the same expected mutations and wild type genotypes as the reference method, with few exceptions that correlated with very low parasitaemia samples. In addition, two cases of heterozygotes that had not been detected by sequencing were found. No correlation was found between the MOI or parasitaemia and the KASP values of the sample. The reproducibility of the technique shows no oscillations between repetitions in any of the three SNPs analysed. Conclusions The KASP assays developed in this study were efficient and versatile for the determination of the Plasmodium genotypes related to ... |
format |
Article in Journal/Newspaper |
author |
Ana Alvarez-Fernandez María J. Bernal Isabel Fradejas Alexandra Martin Ramírez Noor Azian Md Yusuf Marta Lanza Shamilah Hisam Ana Pérez de Ayala José M. Rubio |
author_facet |
Ana Alvarez-Fernandez María J. Bernal Isabel Fradejas Alexandra Martin Ramírez Noor Azian Md Yusuf Marta Lanza Shamilah Hisam Ana Pérez de Ayala José M. Rubio |
author_sort |
Ana Alvarez-Fernandez |
title |
KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum |
title_short |
KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum |
title_full |
KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum |
title_fullStr |
KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum |
title_full_unstemmed |
KASP: a genotyping method to rapid identification of resistance in Plasmodium falciparum |
title_sort |
kasp: a genotyping method to rapid identification of resistance in plasmodium falciparum |
publisher |
BMC |
publishDate |
2021 |
url |
https://doi.org/10.1186/s12936-020-03544-7 https://doaj.org/article/dea477b7b63349e883bcd864bece77ea |
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Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 20, Iss 1, Pp 1-8 (2021) |
op_relation |
https://doi.org/10.1186/s12936-020-03544-7 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-020-03544-7 1475-2875 https://doaj.org/article/dea477b7b63349e883bcd864bece77ea |
op_doi |
https://doi.org/10.1186/s12936-020-03544-7 |
container_title |
Malaria Journal |
container_volume |
20 |
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1 |
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1766347834712391680 |