A simple PCR-SSP method for detection of HLA-B*15:02, *15:13, and *15:21-Reference-Cited by-同舟云学术

A simple PCR-SSP method for detection of HLA-B*15:02, *15:13, and *15:21

Published:2023-02-08 Issue:3 Volume:48 Page:230-233
ISSN:1303-829X
Container-title:Turkish Journal of Biochemistry
language:en
Short-container-title:

Author:

Musa Nurfadhlina¹^ORCID,Govindasamy G. Saravana²^ORCID,Hoong Chew Beng³^ORCID,Annuar Aziati Azwari¹^ORCID,Mohamed Noor Dzul Azri⁴^ORCID,Daud Nur Aizati Athirah¹⁴^ORCID

Affiliation:

1. Human Genome Centre, School of Medical Sciences , Universiti Sains Malaysia , 16150 Kubang Kerian, Kota Bharu , Kelantan , Malaysia

2. Department of Pharmacy , Hospital Segamat , Ministry of Health Malaysia, Segamat , Johor , Malaysia

3. Department of Pharmacy , Hospital Sultanah Bahiyah , Ministry of Health Malaysia, Alor Setar , Kedah , Malaysia

4. Discipline of Clinical Pharmacy, School of Pharmaceutical Sciences , Universiti Sains Malaysia , 11800 USM , Pulau Pinang , Malaysia

Abstract

Abstract Objectives This study aims to develop a simple polymerase chain reaction (PCR) sequence-specific primer method, which will be used to genotype HLA-B*15:02, HLA-B*15:13, and HLA-B*15:21. Methods DNA was extracted from whole blood using a commercial DNA extraction kit. New specific primers were designed. The PCR was optimized for reproducibility and specificity. Parameters investigated included concentrations of MgCl2, primers concentration, and annealing temperature, to produce specific bands of interest. Known DNA samples were selected at random and tested using the PCR method. Results Simultaneously six duplex PCRs were successfully optimized using 1.0 mM MgCl2 and an annealing temperature of 62 °C. The method was also reproducible and specific. The results from the method showed 100% concordance with known DNA samples. Conclusions This study has successfully developed a simple and specific method to simultaneously genotype HLA-B*15:02, HLA-B*15:13, and HLA-B*15:21 using in-house developed PCR-SSP.

Funder

Universiti Sains Malaysia Short Term Grant

Publisher

Walter de Gruyter GmbH

Subject

Biochemistry (medical),Clinical Biochemistry,Molecular Biology,Biochemistry

Link

https://www.degruyter.com/document/doi/10.1515/tjb-2022-0202/pdf

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