Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification
Abstract Background Anopheles gambiae M and S molecular forms, the major malaria vectors in the Afro-tropical region, are ongoing a process of ecological diversification and adaptive lineage splitting, which is affecting malaria transmission and vector control strategies in West Africa. These two in...
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ftdoajarticles:oai:doaj.org/article:0070e3689d654f028f3394ffb936c8bb 2023-05-15T15:18:02+02:00 Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification Pinto João Petrarca Vincenzo Mancini Emiliano Vicente José L Calzetta Maria Caputo Beniamino Santolamazza Federica della Torre Alessandra 2011-08-01T00:00:00Z https://doi.org/10.1186/1475-2875-10-215 https://doaj.org/article/0070e3689d654f028f3394ffb936c8bb EN eng BMC http://www.malariajournal.com/content/10/1/215 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-10-215 1475-2875 https://doaj.org/article/0070e3689d654f028f3394ffb936c8bb Malaria Journal, Vol 10, Iss 1, p 215 (2011) Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2011 ftdoajarticles https://doi.org/10.1186/1475-2875-10-215 2022-12-31T08:15:14Z Abstract Background Anopheles gambiae M and S molecular forms, the major malaria vectors in the Afro-tropical region, are ongoing a process of ecological diversification and adaptive lineage splitting, which is affecting malaria transmission and vector control strategies in West Africa. These two incipient species are defined on the basis of single nucleotide differences in the IGS and ITS regions of multicopy rDNA located on the X-chromosome. A number of PCR and PCR-RFLP approaches based on form-specific SNPs in the IGS region are used for M and S identification. Moreover, a PCR-method to detect the M-specific insertion of a short interspersed transposable element ( SINE200 ) has recently been introduced as an alternative identification approach. However, a large-scale comparative analysis of four widely used PCR or PCR-RFLP genotyping methods for M and S identification was never carried out to evaluate whether they could be used interchangeably, as commonly assumed. Results The genotyping of more than 400 A. gambiae specimens from nine African countries, and the sequencing of the IGS-amplicon of 115 of them, highlighted discrepancies among results obtained by the different approaches due to different kinds of biases, which may result in an overestimation of MS putative hybrids, as follows: i) incorrect match of M and S specific primers used in the allele specific-PCR approach; ii) presence of polymorphisms in the recognition sequence of restriction enzymes used in the PCR-RFLP approaches; iii) incomplete cleavage during the restriction reactions; iv) presence of different copy numbers of M and S-specific IGS-arrays in single individuals in areas of secondary contact between the two forms. Conclusions The results reveal that the PCR and PCR-RFLP approaches most commonly utilized to identify A. gambiae M and S forms are not fully interchangeable as usually assumed, and highlight limits of the actual definition of the two molecular forms, which might not fully correspond to the two A. gambiae incipient species in ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 10 1 215 |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Pinto João Petrarca Vincenzo Mancini Emiliano Vicente José L Calzetta Maria Caputo Beniamino Santolamazza Federica della Torre Alessandra Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Anopheles gambiae M and S molecular forms, the major malaria vectors in the Afro-tropical region, are ongoing a process of ecological diversification and adaptive lineage splitting, which is affecting malaria transmission and vector control strategies in West Africa. These two incipient species are defined on the basis of single nucleotide differences in the IGS and ITS regions of multicopy rDNA located on the X-chromosome. A number of PCR and PCR-RFLP approaches based on form-specific SNPs in the IGS region are used for M and S identification. Moreover, a PCR-method to detect the M-specific insertion of a short interspersed transposable element ( SINE200 ) has recently been introduced as an alternative identification approach. However, a large-scale comparative analysis of four widely used PCR or PCR-RFLP genotyping methods for M and S identification was never carried out to evaluate whether they could be used interchangeably, as commonly assumed. Results The genotyping of more than 400 A. gambiae specimens from nine African countries, and the sequencing of the IGS-amplicon of 115 of them, highlighted discrepancies among results obtained by the different approaches due to different kinds of biases, which may result in an overestimation of MS putative hybrids, as follows: i) incorrect match of M and S specific primers used in the allele specific-PCR approach; ii) presence of polymorphisms in the recognition sequence of restriction enzymes used in the PCR-RFLP approaches; iii) incomplete cleavage during the restriction reactions; iv) presence of different copy numbers of M and S-specific IGS-arrays in single individuals in areas of secondary contact between the two forms. Conclusions The results reveal that the PCR and PCR-RFLP approaches most commonly utilized to identify A. gambiae M and S forms are not fully interchangeable as usually assumed, and highlight limits of the actual definition of the two molecular forms, which might not fully correspond to the two A. gambiae incipient species in ... |
format |
Article in Journal/Newspaper |
author |
Pinto João Petrarca Vincenzo Mancini Emiliano Vicente José L Calzetta Maria Caputo Beniamino Santolamazza Federica della Torre Alessandra |
author_facet |
Pinto João Petrarca Vincenzo Mancini Emiliano Vicente José L Calzetta Maria Caputo Beniamino Santolamazza Federica della Torre Alessandra |
author_sort |
Pinto João |
title |
Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification |
title_short |
Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification |
title_full |
Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification |
title_fullStr |
Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification |
title_full_unstemmed |
Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiae molecular form identification |
title_sort |
comparative analyses reveal discrepancies among results of commonly used methods for anopheles gambiae molecular form identification |
publisher |
BMC |
publishDate |
2011 |
url |
https://doi.org/10.1186/1475-2875-10-215 https://doaj.org/article/0070e3689d654f028f3394ffb936c8bb |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 10, Iss 1, p 215 (2011) |
op_relation |
http://www.malariajournal.com/content/10/1/215 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-10-215 1475-2875 https://doaj.org/article/0070e3689d654f028f3394ffb936c8bb |
op_doi |
https://doi.org/10.1186/1475-2875-10-215 |
container_title |
Malaria Journal |
container_volume |
10 |
container_issue |
1 |
container_start_page |
215 |
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1766348267136745472 |