Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses

We used quantitative real-time polymerase chain reaction (qPCR) and high resolution melting (HRM) analyses for the detection of the common α-thalassemia (α-thal) genotypes in 40 northern Thai Hb H (β4) patients. The α 0 -thal [- SEA (Southeast Asian) deletion] was detected by a multiplex gap real-ti...

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Main Authors: Teerapat Seeratanachot, Torpong Sanguansermsri, Dawan Shimbhu
Format: Journal
Published: 2018
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/47499
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-474992018-04-25T08:40:45Z Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses Teerapat Seeratanachot Torpong Sanguansermsri Dawan Shimbhu We used quantitative real-time polymerase chain reaction (qPCR) and high resolution melting (HRM) analyses for the detection of the common α-thalassemia (α-thal) genotypes in 40 northern Thai Hb H (β4) patients. The α 0 -thal [- SEA (Southeast Asian) deletion] was detected by a multiplex gap real-time PCR. To determine the α + -thal, three primer pairs were designed. The A-primer pair was used to amplify the 3' terminal DNA sequences of the HBA2 gene, and the B-primer pair amplified the 5' flanking region of the HBA1 gene. The C-primer pair amplified the 3' terminal DNA sequences of the HBA1 gene and was used as an internal control. The-α 4.2 (leftward) and-α 3.7 (rightward) deletions were determined by monitoring the absence of PCR product(s). The Hb H patients who had a negative PCR result for the A-primer pair but positive for the B-and C-primer pairs carried the-α 4.2 deletion, while the-α 3.7 deletion carriers were negative for the A-and B-primer pairs. In the case of Hb H with Hb Constant Spring (Hb CS, α142, Term→Gln; HBA2: c.427T > C), all primer pairs were positive, HRM analysis of the PCR product of the A-primer pair was introduced to analyze the Hb CS gene. It can distinguish clearly between normal and Hb CS genotypes. The applied methods for the determination of the α 0 -and α + -thal genotypes revealed the results to be as accurate as conventional gap-PCR and the direct DNA sequencing methods but resulted in a much simpler and faster procedure. © Informa Healthcare USA, Inc. 2018-04-25T08:40:45Z 2018-04-25T08:40:45Z 2013-11-13 Journal 1532432X 03630269 2-s2.0-84887215064 10.3109/03630269.2013.828228 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84887215064&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/47499
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
description We used quantitative real-time polymerase chain reaction (qPCR) and high resolution melting (HRM) analyses for the detection of the common α-thalassemia (α-thal) genotypes in 40 northern Thai Hb H (β4) patients. The α 0 -thal [- SEA (Southeast Asian) deletion] was detected by a multiplex gap real-time PCR. To determine the α + -thal, three primer pairs were designed. The A-primer pair was used to amplify the 3' terminal DNA sequences of the HBA2 gene, and the B-primer pair amplified the 5' flanking region of the HBA1 gene. The C-primer pair amplified the 3' terminal DNA sequences of the HBA1 gene and was used as an internal control. The-α 4.2 (leftward) and-α 3.7 (rightward) deletions were determined by monitoring the absence of PCR product(s). The Hb H patients who had a negative PCR result for the A-primer pair but positive for the B-and C-primer pairs carried the-α 4.2 deletion, while the-α 3.7 deletion carriers were negative for the A-and B-primer pairs. In the case of Hb H with Hb Constant Spring (Hb CS, α142, Term→Gln; HBA2: c.427T > C), all primer pairs were positive, HRM analysis of the PCR product of the A-primer pair was introduced to analyze the Hb CS gene. It can distinguish clearly between normal and Hb CS genotypes. The applied methods for the determination of the α 0 -and α + -thal genotypes revealed the results to be as accurate as conventional gap-PCR and the direct DNA sequencing methods but resulted in a much simpler and faster procedure. © Informa Healthcare USA, Inc.
format Journal
author Teerapat Seeratanachot
Torpong Sanguansermsri
Dawan Shimbhu
spellingShingle Teerapat Seeratanachot
Torpong Sanguansermsri
Dawan Shimbhu
Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses
author_facet Teerapat Seeratanachot
Torpong Sanguansermsri
Dawan Shimbhu
author_sort Teerapat Seeratanachot
title Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses
title_short Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses
title_full Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses
title_fullStr Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses
title_full_unstemmed Detection of Hb H disease genotypes common in Northern Thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses
title_sort detection of hb h disease genotypes common in northern thailand by quantitative real-time polymerase chain reaction and high resolution melting analyses
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84887215064&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/47499
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