IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis
Abstract Background Polymerase chain reactions to distinguish single-nucleotide polymorphisms are commonly used for mosquito identification and identifying insecticide resistance alleles. However, the existing methods used for primer design often result in analyses that are not robust or require add...
Published in: | Malaria Journal |
---|---|
Main Authors: | , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
BMC
2006
|
Subjects: | |
Online Access: | https://doi.org/10.1186/1475-2875-5-125 https://doaj.org/article/a793d3e477f34d46ad87ecd71b0e0e9c |
id |
ftdoajarticles:oai:doaj.org/article:a793d3e477f34d46ad87ecd71b0e0e9c |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:a793d3e477f34d46ad87ecd71b0e0e9c 2023-05-15T15:08:05+02:00 IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis Howell Paul I Wilkins Elien E Benedict Mark Q 2006-12-01T00:00:00Z https://doi.org/10.1186/1475-2875-5-125 https://doaj.org/article/a793d3e477f34d46ad87ecd71b0e0e9c EN eng BMC http://www.malariajournal.com/content/5/1/125 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-5-125 1475-2875 https://doaj.org/article/a793d3e477f34d46ad87ecd71b0e0e9c Malaria Journal, Vol 5, Iss 1, p 125 (2006) Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2006 ftdoajarticles https://doi.org/10.1186/1475-2875-5-125 2022-12-31T09:11:03Z Abstract Background Polymerase chain reactions to distinguish single-nucleotide polymorphisms are commonly used for mosquito identification and identifying insecticide resistance alleles. However, the existing methods used for primer design often result in analyses that are not robust or require additional steps. Methods Utilizing oligonucleotides that are unique in having an intentional mismatch to both templates three bases from the SNP at the 3-prime end, three new PCR assays that distinguish SNP targets using standard gel electrophoresis of undigested DNA fragments were developed and tested. These were applied to: (1) an alternative ribosomal DNA PCR assay to distinguish five members of the Anopheles gambiae complex; (2) detection of the Mopti and Savanna rDNA types; and (3) an assay to distinguish resistance to dieldrin ( Rdl ) alleles in Anopheles arabiensis . Results Reproducible specific amplification of the target alleles was observed in all three assays. The results were consistent with existing analyses but proved simpler and the results more distinct in our hands. Conclusion The simplicity and effectiveness of the method should be utilized in these and other PCR analyses to increase their specificity and simplicity. These results have the potential to be extended not only to mosquito analyses but also to parasite and human polymorphisms. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 5 1 125 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Howell Paul I Wilkins Elien E Benedict Mark Q IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Polymerase chain reactions to distinguish single-nucleotide polymorphisms are commonly used for mosquito identification and identifying insecticide resistance alleles. However, the existing methods used for primer design often result in analyses that are not robust or require additional steps. Methods Utilizing oligonucleotides that are unique in having an intentional mismatch to both templates three bases from the SNP at the 3-prime end, three new PCR assays that distinguish SNP targets using standard gel electrophoresis of undigested DNA fragments were developed and tested. These were applied to: (1) an alternative ribosomal DNA PCR assay to distinguish five members of the Anopheles gambiae complex; (2) detection of the Mopti and Savanna rDNA types; and (3) an assay to distinguish resistance to dieldrin ( Rdl ) alleles in Anopheles arabiensis . Results Reproducible specific amplification of the target alleles was observed in all three assays. The results were consistent with existing analyses but proved simpler and the results more distinct in our hands. Conclusion The simplicity and effectiveness of the method should be utilized in these and other PCR analyses to increase their specificity and simplicity. These results have the potential to be extended not only to mosquito analyses but also to parasite and human polymorphisms. |
format |
Article in Journal/Newspaper |
author |
Howell Paul I Wilkins Elien E Benedict Mark Q |
author_facet |
Howell Paul I Wilkins Elien E Benedict Mark Q |
author_sort |
Howell Paul I |
title |
IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis |
title_short |
IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis |
title_full |
IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis |
title_fullStr |
IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis |
title_full_unstemmed |
IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis |
title_sort |
imp pcr primers detect single nucleotide polymorphisms for anopheles gambiae species identification, mopti and savanna rdna types, and resistance to dieldrin in anopheles arabiensis |
publisher |
BMC |
publishDate |
2006 |
url |
https://doi.org/10.1186/1475-2875-5-125 https://doaj.org/article/a793d3e477f34d46ad87ecd71b0e0e9c |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 5, Iss 1, p 125 (2006) |
op_relation |
http://www.malariajournal.com/content/5/1/125 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-5-125 1475-2875 https://doaj.org/article/a793d3e477f34d46ad87ecd71b0e0e9c |
op_doi |
https://doi.org/10.1186/1475-2875-5-125 |
container_title |
Malaria Journal |
container_volume |
5 |
container_issue |
1 |
container_start_page |
125 |
_version_ |
1766339516797288448 |