HLA-B*15: 02 screening in epileptic patients using a high resolution melting-real time PCR (HRM-QPCR) method

Zam Zureena Mohd Rani, Nor Azian Abdul Murad, Sue Mian Then, Suthashini Panja Bernam, Asmaa Abdullah, Saberi Saimun, Sri Noraima Othman, Raymond Azman Ali, A. Rahman A. Jamal

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background: The HLA-B*15:02 polymorphism in epileptic patients is known to be associated with carbamazepine-induced Stevens-Johnson syndrome (SJS). The prevalence of HLA-B*15:02 polymorphism seemed to be ethnic-specific with a higher frequency of HLA-B*15:02 in Asian compared to the Europeans. This study was performed to determine the frequency of the HLA-B*15:02 polymorphism in epileptic patients at the Chancellor Tuanku Muhriz Hospital-UKM Medical Centre (HCTM-UKMMC) using high resolution melting-real time PCR (HRM-QPCR) method. Methods: We performed a fast and effective in-house high resolution melting-real time polymerase chain reaction method and compared it with the conventional multiplex-PCR method. The specificity and sensitivity of each test were also determined using DNA from saliva. Results: Using the conventional multiplex-PCR approach for screening, 25 out of 64 (39.1%) epileptic patients were positive for HLA-B*15:02. However, using the HRM-QPCR technique, 24/64 (37.5%) of the patients were positive. The one patient who tested positive by the multiplex-PCR but negative using the HRM-QPCR turned out to be negative by DNA sequencing. The HRM-QPCR and DNA sequencing showed 100% sensitivity and specificity. The multiplex-PCR showed 100% sensitivity and 98.4% specificity compared to both HRM-QPCR and DNA sequencing. The HRM-QPCR is also more cost-effective (<$16.40 USD/test) and less time-consuming when compared to the multiplex-PCR ($25.15 USD/test). Conclusion: Our result suggested that multiplex PCR, HRM-QPCR and Sanger sequencing can be used for detection of HLA-B*15:02. However, a qualitative method such as multiplex PCR should be confirmed with other quantitative methods such as HRM-QPCR and Sanger sequencing.

Original languageEnglish
Pages (from-to)137-144
Number of pages8
JournalNeurology Asia
Volume23
Issue number2
Publication statusPublished - 1 Jun 2018

Fingerprint

HLA-B Antigens
Multiplex Polymerase Chain Reaction
Freezing
Real-Time Polymerase Chain Reaction
DNA Sequence Analysis
Sensitivity and Specificity
Stevens-Johnson Syndrome
Carbamazepine
Saliva
Costs and Cost Analysis
DNA

Keywords

  • Carbamazepine-induced Steven Johnson syndrome
  • DNA sequencing
  • Epilepsy
  • High resolution melting-real time polymerase chain reaction (HRM-QPCR)
  • Multiplex-polymerase chain reaction

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology

Cite this

HLA-B*15 : 02 screening in epileptic patients using a high resolution melting-real time PCR (HRM-QPCR) method. / Mohd Rani, Zam Zureena; Abdul Murad, Nor Azian; Then, Sue Mian; Bernam, Suthashini Panja; Abdullah, Asmaa; Saimun, Saberi; Othman, Sri Noraima; Ali, Raymond Azman; A. Jamal, A. Rahman.

In: Neurology Asia, Vol. 23, No. 2, 01.06.2018, p. 137-144.

Research output: Contribution to journalArticle

Mohd Rani, ZZ, Abdul Murad, NA, Then, SM, Bernam, SP, Abdullah, A, Saimun, S, Othman, SN, Ali, RA & A. Jamal, AR 2018, 'HLA-B*15: 02 screening in epileptic patients using a high resolution melting-real time PCR (HRM-QPCR) method', Neurology Asia, vol. 23, no. 2, pp. 137-144.
Mohd Rani, Zam Zureena ; Abdul Murad, Nor Azian ; Then, Sue Mian ; Bernam, Suthashini Panja ; Abdullah, Asmaa ; Saimun, Saberi ; Othman, Sri Noraima ; Ali, Raymond Azman ; A. Jamal, A. Rahman. / HLA-B*15 : 02 screening in epileptic patients using a high resolution melting-real time PCR (HRM-QPCR) method. In: Neurology Asia. 2018 ; Vol. 23, No. 2. pp. 137-144.
@article{29aa8adee290418b9af931316a637862,
title = "HLA-B*15: 02 screening in epileptic patients using a high resolution melting-real time PCR (HRM-QPCR) method",
abstract = "Background: The HLA-B*15:02 polymorphism in epileptic patients is known to be associated with carbamazepine-induced Stevens-Johnson syndrome (SJS). The prevalence of HLA-B*15:02 polymorphism seemed to be ethnic-specific with a higher frequency of HLA-B*15:02 in Asian compared to the Europeans. This study was performed to determine the frequency of the HLA-B*15:02 polymorphism in epileptic patients at the Chancellor Tuanku Muhriz Hospital-UKM Medical Centre (HCTM-UKMMC) using high resolution melting-real time PCR (HRM-QPCR) method. Methods: We performed a fast and effective in-house high resolution melting-real time polymerase chain reaction method and compared it with the conventional multiplex-PCR method. The specificity and sensitivity of each test were also determined using DNA from saliva. Results: Using the conventional multiplex-PCR approach for screening, 25 out of 64 (39.1{\%}) epileptic patients were positive for HLA-B*15:02. However, using the HRM-QPCR technique, 24/64 (37.5{\%}) of the patients were positive. The one patient who tested positive by the multiplex-PCR but negative using the HRM-QPCR turned out to be negative by DNA sequencing. The HRM-QPCR and DNA sequencing showed 100{\%} sensitivity and specificity. The multiplex-PCR showed 100{\%} sensitivity and 98.4{\%} specificity compared to both HRM-QPCR and DNA sequencing. The HRM-QPCR is also more cost-effective (<$16.40 USD/test) and less time-consuming when compared to the multiplex-PCR ($25.15 USD/test). Conclusion: Our result suggested that multiplex PCR, HRM-QPCR and Sanger sequencing can be used for detection of HLA-B*15:02. However, a qualitative method such as multiplex PCR should be confirmed with other quantitative methods such as HRM-QPCR and Sanger sequencing.",
keywords = "Carbamazepine-induced Steven Johnson syndrome, DNA sequencing, Epilepsy, High resolution melting-real time polymerase chain reaction (HRM-QPCR), Multiplex-polymerase chain reaction",
author = "{Mohd Rani}, {Zam Zureena} and {Abdul Murad}, {Nor Azian} and Then, {Sue Mian} and Bernam, {Suthashini Panja} and Asmaa Abdullah and Saberi Saimun and Othman, {Sri Noraima} and Ali, {Raymond Azman} and {A. Jamal}, {A. Rahman}",
year = "2018",
month = "6",
day = "1",
language = "English",
volume = "23",
pages = "137--144",
journal = "Neurology Asia",
issn = "1823-6138",
publisher = "ASEAN Neurological Association",
number = "2",

}

TY - JOUR

T1 - HLA-B*15

T2 - 02 screening in epileptic patients using a high resolution melting-real time PCR (HRM-QPCR) method

AU - Mohd Rani, Zam Zureena

AU - Abdul Murad, Nor Azian

AU - Then, Sue Mian

AU - Bernam, Suthashini Panja

AU - Abdullah, Asmaa

AU - Saimun, Saberi

AU - Othman, Sri Noraima

AU - Ali, Raymond Azman

AU - A. Jamal, A. Rahman

PY - 2018/6/1

Y1 - 2018/6/1

N2 - Background: The HLA-B*15:02 polymorphism in epileptic patients is known to be associated with carbamazepine-induced Stevens-Johnson syndrome (SJS). The prevalence of HLA-B*15:02 polymorphism seemed to be ethnic-specific with a higher frequency of HLA-B*15:02 in Asian compared to the Europeans. This study was performed to determine the frequency of the HLA-B*15:02 polymorphism in epileptic patients at the Chancellor Tuanku Muhriz Hospital-UKM Medical Centre (HCTM-UKMMC) using high resolution melting-real time PCR (HRM-QPCR) method. Methods: We performed a fast and effective in-house high resolution melting-real time polymerase chain reaction method and compared it with the conventional multiplex-PCR method. The specificity and sensitivity of each test were also determined using DNA from saliva. Results: Using the conventional multiplex-PCR approach for screening, 25 out of 64 (39.1%) epileptic patients were positive for HLA-B*15:02. However, using the HRM-QPCR technique, 24/64 (37.5%) of the patients were positive. The one patient who tested positive by the multiplex-PCR but negative using the HRM-QPCR turned out to be negative by DNA sequencing. The HRM-QPCR and DNA sequencing showed 100% sensitivity and specificity. The multiplex-PCR showed 100% sensitivity and 98.4% specificity compared to both HRM-QPCR and DNA sequencing. The HRM-QPCR is also more cost-effective (<$16.40 USD/test) and less time-consuming when compared to the multiplex-PCR ($25.15 USD/test). Conclusion: Our result suggested that multiplex PCR, HRM-QPCR and Sanger sequencing can be used for detection of HLA-B*15:02. However, a qualitative method such as multiplex PCR should be confirmed with other quantitative methods such as HRM-QPCR and Sanger sequencing.

AB - Background: The HLA-B*15:02 polymorphism in epileptic patients is known to be associated with carbamazepine-induced Stevens-Johnson syndrome (SJS). The prevalence of HLA-B*15:02 polymorphism seemed to be ethnic-specific with a higher frequency of HLA-B*15:02 in Asian compared to the Europeans. This study was performed to determine the frequency of the HLA-B*15:02 polymorphism in epileptic patients at the Chancellor Tuanku Muhriz Hospital-UKM Medical Centre (HCTM-UKMMC) using high resolution melting-real time PCR (HRM-QPCR) method. Methods: We performed a fast and effective in-house high resolution melting-real time polymerase chain reaction method and compared it with the conventional multiplex-PCR method. The specificity and sensitivity of each test were also determined using DNA from saliva. Results: Using the conventional multiplex-PCR approach for screening, 25 out of 64 (39.1%) epileptic patients were positive for HLA-B*15:02. However, using the HRM-QPCR technique, 24/64 (37.5%) of the patients were positive. The one patient who tested positive by the multiplex-PCR but negative using the HRM-QPCR turned out to be negative by DNA sequencing. The HRM-QPCR and DNA sequencing showed 100% sensitivity and specificity. The multiplex-PCR showed 100% sensitivity and 98.4% specificity compared to both HRM-QPCR and DNA sequencing. The HRM-QPCR is also more cost-effective (<$16.40 USD/test) and less time-consuming when compared to the multiplex-PCR ($25.15 USD/test). Conclusion: Our result suggested that multiplex PCR, HRM-QPCR and Sanger sequencing can be used for detection of HLA-B*15:02. However, a qualitative method such as multiplex PCR should be confirmed with other quantitative methods such as HRM-QPCR and Sanger sequencing.

KW - Carbamazepine-induced Steven Johnson syndrome

KW - DNA sequencing

KW - Epilepsy

KW - High resolution melting-real time polymerase chain reaction (HRM-QPCR)

KW - Multiplex-polymerase chain reaction

UR - http://www.scopus.com/inward/record.url?scp=85049451132&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85049451132&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:85049451132

VL - 23

SP - 137

EP - 144

JO - Neurology Asia

JF - Neurology Asia

SN - 1823-6138

IS - 2

ER -