Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus
Objective: To develop the multiplex PCR method based on the internal transcribed spacer 2 to discriminate the intestinal trematodes, Centrocestus caninus (C. caninus), and Stellantchasmus falcatus (S. falcatus). Methods: Four species of heterophyid trematodes including C. caninus, S. falcatus, Haplo...
Published in: | Asian Pacific Journal of Tropical Biomedicine |
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Wolters Kluwer Medknow Publications
2017
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ftdoajarticles:oai:doaj.org/article:4e38ed559c8b42e793e418397664b12f 2023-05-15T15:08:48+02:00 Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus Thapana Chontananarth 2017-02-01T00:00:00Z https://doi.org/10.1016/j.apjtb.2016.11.018 https://doaj.org/article/4e38ed559c8b42e793e418397664b12f EN eng Wolters Kluwer Medknow Publications http://www.sciencedirect.com/science/article/pii/S2221169116302568 https://doaj.org/toc/2221-1691 2221-1691 doi:10.1016/j.apjtb.2016.11.018 https://doaj.org/article/4e38ed559c8b42e793e418397664b12f Asian Pacific Journal of Tropical Biomedicine, Vol 7, Iss 2, Pp 103-106 (2017) Multiplex PCR Centrocestus caninus Stellantchasmus falcatus ITS2 Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 article 2017 ftdoajarticles https://doi.org/10.1016/j.apjtb.2016.11.018 2022-12-31T15:41:01Z Objective: To develop the multiplex PCR method based on the internal transcribed spacer 2 to discriminate the intestinal trematodes, Centrocestus caninus (C. caninus), and Stellantchasmus falcatus (S. falcatus). Methods: Four species of heterophyid trematodes including C. caninus, S. falcatus, Haplorchis taichui and Haplorchoides sp. were amplified and the specific primer was designed based on the internal transcribed spacer 2 region. Two specific primers were used to validate the optimized PCR conditions: the specificity test and the sensitivity test. Results: Both of these specific primers confirmed the specificity through multiplex PCR reaction which generated both PCR products (231 and 137 bp) in the mixed DNA template of C. caninus and S. falcatus with no cross-reaction with other heterophyid trematodes. The optimum annealing temperature of both primers was 54–59 °C. The sensitivity test used the two-fold serial dilution DNA template, which was concentrated between 10 and 0.3125 ng/μL. The lowest concentration of the DNA template of this multiplex PCR was 2.5 ng/μL. Conclusions: The technique described here proved to be a species-specific technique and was found to be a rapid method for the diagnosis of C. caninus and S. falcatus in terms of the larval and adult stages in intermediate and/or definitive hosts in the endemic area. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Asian Pacific Journal of Tropical Biomedicine 7 2 103 106 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Multiplex PCR Centrocestus caninus Stellantchasmus falcatus ITS2 Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 |
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Multiplex PCR Centrocestus caninus Stellantchasmus falcatus ITS2 Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 Thapana Chontananarth Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus |
topic_facet |
Multiplex PCR Centrocestus caninus Stellantchasmus falcatus ITS2 Arctic medicine. Tropical medicine RC955-962 Biology (General) QH301-705.5 |
description |
Objective: To develop the multiplex PCR method based on the internal transcribed spacer 2 to discriminate the intestinal trematodes, Centrocestus caninus (C. caninus), and Stellantchasmus falcatus (S. falcatus). Methods: Four species of heterophyid trematodes including C. caninus, S. falcatus, Haplorchis taichui and Haplorchoides sp. were amplified and the specific primer was designed based on the internal transcribed spacer 2 region. Two specific primers were used to validate the optimized PCR conditions: the specificity test and the sensitivity test. Results: Both of these specific primers confirmed the specificity through multiplex PCR reaction which generated both PCR products (231 and 137 bp) in the mixed DNA template of C. caninus and S. falcatus with no cross-reaction with other heterophyid trematodes. The optimum annealing temperature of both primers was 54–59 °C. The sensitivity test used the two-fold serial dilution DNA template, which was concentrated between 10 and 0.3125 ng/μL. The lowest concentration of the DNA template of this multiplex PCR was 2.5 ng/μL. Conclusions: The technique described here proved to be a species-specific technique and was found to be a rapid method for the diagnosis of C. caninus and S. falcatus in terms of the larval and adult stages in intermediate and/or definitive hosts in the endemic area. |
format |
Article in Journal/Newspaper |
author |
Thapana Chontananarth |
author_facet |
Thapana Chontananarth |
author_sort |
Thapana Chontananarth |
title |
Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus |
title_short |
Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus |
title_full |
Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus |
title_fullStr |
Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus |
title_full_unstemmed |
Multiplex PCR assay for discrimination of Centrocestus caninus and Stellantchasmus falcatus |
title_sort |
multiplex pcr assay for discrimination of centrocestus caninus and stellantchasmus falcatus |
publisher |
Wolters Kluwer Medknow Publications |
publishDate |
2017 |
url |
https://doi.org/10.1016/j.apjtb.2016.11.018 https://doaj.org/article/4e38ed559c8b42e793e418397664b12f |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Asian Pacific Journal of Tropical Biomedicine, Vol 7, Iss 2, Pp 103-106 (2017) |
op_relation |
http://www.sciencedirect.com/science/article/pii/S2221169116302568 https://doaj.org/toc/2221-1691 2221-1691 doi:10.1016/j.apjtb.2016.11.018 https://doaj.org/article/4e38ed559c8b42e793e418397664b12f |
op_doi |
https://doi.org/10.1016/j.apjtb.2016.11.018 |
container_title |
Asian Pacific Journal of Tropical Biomedicine |
container_volume |
7 |
container_issue |
2 |
container_start_page |
103 |
op_container_end_page |
106 |
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1766340087130357760 |