A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay
This article reports a rapid and effective method for the extraction and purification of genomic DNA (gDNA) from individual first-stage larvae (L 1 ) of elaphostrongyline nematodes that had been stored frozen or fixed in 95% ethanol for 1 to 5 years. The method was highly effective for L 1 s of all...
Published in: | Journal of Veterinary Diagnostic Investigation |
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Online Access: | http://dx.doi.org/10.1177/104063870501700612 http://journals.sagepub.com/doi/pdf/10.1177/104063870501700612 |
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crsagepubl:10.1177/104063870501700612 2023-05-15T17:22:21+02:00 A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay Chilton, Neil B. Huby-Chilton, Florence Lankester, Murray W. Gajadhar, Alvin A. 2005 http://dx.doi.org/10.1177/104063870501700612 http://journals.sagepub.com/doi/pdf/10.1177/104063870501700612 en eng SAGE Publications http://journals.sagepub.com/page/policies/text-and-data-mining-license Journal of Veterinary Diagnostic Investigation volume 17, issue 6, page 585-588 ISSN 1040-6387 1943-4936 General Veterinary journal-article 2005 crsagepubl https://doi.org/10.1177/104063870501700612 2022-04-14T04:45:52Z This article reports a rapid and effective method for the extraction and purification of genomic DNA (gDNA) from individual first-stage larvae (L 1 ) of elaphostrongyline nematodes that had been stored frozen or fixed in 95% ethanol for 1 to 5 years. The method was highly effective for L 1 s of all 6 species of elaphostrongylines, based on polymerase chain reaction (PCR) amplification of a partial fragment of the first internal transcribed spacer (ITS-1) of the ribosomal DNA. Differences were detected in the sizes of partial ITS-1 amplicons between the 2 elaphostrongyline genera, Elaphostrongylus and Parelaphostrongylus. The reliability of the ITS-1 PCR assay was tested by using L 1 s of unknown identity from Newfoundland and Labrador, Canada. The ability to consistently isolate gDNA from individual L 1 s, together with a simple PCR-based method to distinguish between Parelaphostrongylus and Elaphostrongylus, have important implications for diagnostic testing and for conducting epizootiological studies on these parasites of veterinary importance. Article in Journal/Newspaper Newfoundland SAGE Publications (via Crossref) Canada Newfoundland Journal of Veterinary Diagnostic Investigation 17 6 585 588 |
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Open Polar |
collection |
SAGE Publications (via Crossref) |
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crsagepubl |
language |
English |
topic |
General Veterinary |
spellingShingle |
General Veterinary Chilton, Neil B. Huby-Chilton, Florence Lankester, Murray W. Gajadhar, Alvin A. A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay |
topic_facet |
General Veterinary |
description |
This article reports a rapid and effective method for the extraction and purification of genomic DNA (gDNA) from individual first-stage larvae (L 1 ) of elaphostrongyline nematodes that had been stored frozen or fixed in 95% ethanol for 1 to 5 years. The method was highly effective for L 1 s of all 6 species of elaphostrongylines, based on polymerase chain reaction (PCR) amplification of a partial fragment of the first internal transcribed spacer (ITS-1) of the ribosomal DNA. Differences were detected in the sizes of partial ITS-1 amplicons between the 2 elaphostrongyline genera, Elaphostrongylus and Parelaphostrongylus. The reliability of the ITS-1 PCR assay was tested by using L 1 s of unknown identity from Newfoundland and Labrador, Canada. The ability to consistently isolate gDNA from individual L 1 s, together with a simple PCR-based method to distinguish between Parelaphostrongylus and Elaphostrongylus, have important implications for diagnostic testing and for conducting epizootiological studies on these parasites of veterinary importance. |
format |
Article in Journal/Newspaper |
author |
Chilton, Neil B. Huby-Chilton, Florence Lankester, Murray W. Gajadhar, Alvin A. |
author_facet |
Chilton, Neil B. Huby-Chilton, Florence Lankester, Murray W. Gajadhar, Alvin A. |
author_sort |
Chilton, Neil B. |
title |
A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay |
title_short |
A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay |
title_full |
A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay |
title_fullStr |
A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay |
title_full_unstemmed |
A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay |
title_sort |
method for extracting genomic dna from individual elaphostrongyline (nematoda: protostrongylidae) larvae and differentiation of elaphostrongylus spp. from parelaphostrongylus spp. by pcr assay |
publisher |
SAGE Publications |
publishDate |
2005 |
url |
http://dx.doi.org/10.1177/104063870501700612 http://journals.sagepub.com/doi/pdf/10.1177/104063870501700612 |
geographic |
Canada Newfoundland |
geographic_facet |
Canada Newfoundland |
genre |
Newfoundland |
genre_facet |
Newfoundland |
op_source |
Journal of Veterinary Diagnostic Investigation volume 17, issue 6, page 585-588 ISSN 1040-6387 1943-4936 |
op_rights |
http://journals.sagepub.com/page/policies/text-and-data-mining-license |
op_doi |
https://doi.org/10.1177/104063870501700612 |
container_title |
Journal of Veterinary Diagnostic Investigation |
container_volume |
17 |
container_issue |
6 |
container_start_page |
585 |
op_container_end_page |
588 |
_version_ |
1766108955057061888 |