Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection

Specific primers to determine the presence of an intestinal fluke, Haplorchis taichui, were investigated using the high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR, and 18 arbitrary primers (Operon Technologies), to generate different polymorphic DNA profiles. Thirteen kind...

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Main Authors: Chalobol Wongsawad, Pheravut Wongsawad, Jong Yil Chai, Somboon Anuntalabhochai
Format: Journal
Published: 2018
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/49132
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-491322018-08-16T02:10:39Z Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection Chalobol Wongsawad Pheravut Wongsawad Jong Yil Chai Somboon Anuntalabhochai Immunology and Microbiology Specific primers to determine the presence of an intestinal fluke, Haplorchis taichui, were investigated using the high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR, and 18 arbitrary primers (Operon Technologies), to generate different polymorphic DNA profiles. Thirteen kinds of parasites were used to compare fingerprints. A 256 bp HAT-RAPD marker, generated from the OPP-11 primer, was found to be H. taichui-specific, and this marker was cloned, transformed, and sequenced. From the sequence data, a pair of primers were designed with Genetyx-MAC ver.11 and indicated as: Hap-t F 5′-GGC CAA CGC AAT CGT CAT CC-3′ and Hap-t R 5′-GCG TCG GGT TTC AGA CAT GG-3′. These specific primers were tested for efficacy and specificity by amplifying them with all 13 parasites DNAs in PCR reaction. A 256 bp amplicon was generated, which was shown to have a positive result, only for H. taichui DNA. It revealed no cross-reaction with any of the other tested parasite species. The minimum DNA template, needed for detection by PCR, was 0.1 picogram (pg). The successful development of H. taichui-specific primers is expected to be beneficial for epidemiological studies and for prevention and control of these parasitic infections. © 2009 Elsevier Inc. All rights reserved. 2018-08-16T02:10:39Z 2018-08-16T02:10:39Z 2009-10-01 Journal 10902449 00144894 2-s2.0-68949169193 10.1016/j.exppara.2009.06.016 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=68949169193&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/49132
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Immunology and Microbiology
spellingShingle Immunology and Microbiology
Chalobol Wongsawad
Pheravut Wongsawad
Jong Yil Chai
Somboon Anuntalabhochai
Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection
description Specific primers to determine the presence of an intestinal fluke, Haplorchis taichui, were investigated using the high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR, and 18 arbitrary primers (Operon Technologies), to generate different polymorphic DNA profiles. Thirteen kinds of parasites were used to compare fingerprints. A 256 bp HAT-RAPD marker, generated from the OPP-11 primer, was found to be H. taichui-specific, and this marker was cloned, transformed, and sequenced. From the sequence data, a pair of primers were designed with Genetyx-MAC ver.11 and indicated as: Hap-t F 5′-GGC CAA CGC AAT CGT CAT CC-3′ and Hap-t R 5′-GCG TCG GGT TTC AGA CAT GG-3′. These specific primers were tested for efficacy and specificity by amplifying them with all 13 parasites DNAs in PCR reaction. A 256 bp amplicon was generated, which was shown to have a positive result, only for H. taichui DNA. It revealed no cross-reaction with any of the other tested parasite species. The minimum DNA template, needed for detection by PCR, was 0.1 picogram (pg). The successful development of H. taichui-specific primers is expected to be beneficial for epidemiological studies and for prevention and control of these parasitic infections. © 2009 Elsevier Inc. All rights reserved.
format Journal
author Chalobol Wongsawad
Pheravut Wongsawad
Jong Yil Chai
Somboon Anuntalabhochai
author_facet Chalobol Wongsawad
Pheravut Wongsawad
Jong Yil Chai
Somboon Anuntalabhochai
author_sort Chalobol Wongsawad
title Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection
title_short Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection
title_full Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection
title_fullStr Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection
title_full_unstemmed Haplorchis taichui, Witenberg, 1930: Development of a HAT-RAPD marker for the detection of minute intestinal fluke infection
title_sort haplorchis taichui, witenberg, 1930: development of a hat-rapd marker for the detection of minute intestinal fluke infection
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=68949169193&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/49132
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