COMPARISON OF MUTAGEL HFE AND POLYMERASE CHAIN REACTIONRESTRICTION FRAGMENT LENGTH POLYMORPHISM (PCR-RFLP) FOR THE INVESTIGATION OF C282Y AND H63D POINT MUTATION IN HFE GENE

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ZUJAJA HINA HAROON
HAROON JAVAID
ROSS WILLIAMS
YOUSAF KAZMI

Abstract

OBJECTIVE: To compare MutaGel HFE (commercial kit based on ARMSPCR)with in-house designed polymerase chain reaction-restrictionfragment length polymorphism (PCR-RFLP) techniques for the diagnosisof C282Y and H63D point mutations in HFE-gene, and to recommendthe feasibility of each method for individual laboratories according totheir requirements.METHODOLOGY: This quasi–experimental study was conducted atdepartment of forensic and biomedical sciences, university of LincolnUK from July 2006 to January 2007. Twenty-two samples were analyzedby two different molecular diagnostic techniques (ARMS & RFLP) forthe same genetic disorder (C282Y and H63D mutation in HFE-gene).Blood samples were procured from healthy volunteers from differentgeographical areas. Two controls (one positive and one negative) wererun with each batch. ARMS-PCR was carried out by using Mutagel HFEcommercial kit by immunodiagnostik whereas PCR-RFLP was performedas per UK Haemochromatosis consortium directives.RESULTS: Out of 22 samples for C282Y mutation; 20 showed normalgenotype, one was heterozygous, none was homozygous and 1 showedsample contamination. For H63D mutation, 8 showed normal genotype,one was homozygous, 12 were heterozygous while one sample showedcontamination.There was no statistically significant difference in diagnostic sensitivityand specificity of both groups. Each method had its unique performancecharacteristics that could be utilized by different laboratories dependingon their requirements.CONCLUSION: Both the techniques were equally good for diagnosis ofhaemochromatosis but RFLP semed more suitable for a laboratory withlow workload whereas MutaGel HFE seemed more appropriate for highworkload, less manpower and short turn-around time.KEYWORDS: RFLP (Restriction Fragment Length Polymorphism), PCR(Polymerase Chain Reaction), ARMS (Amplification Refractory MutationSystem), HFE gene mutation, Haemochromatosis.

Article Details

How to Cite
HAROON, Z. H., H. JAVAID, R. WILLIAMS, and Y. KAZMI. “COMPARISON OF MUTAGEL HFE AND POLYMERASE CHAIN REACTIONRESTRICTION FRAGMENT LENGTH POLYMORPHISM (PCR-RFLP) FOR THE INVESTIGATION OF C282Y AND H63D POINT MUTATION IN HFE GENE”. KHYBER MEDICAL UNIVERSITY JOURNAL, vol. 5, no. 2, June 2013, https://www.kmuj.kmu.edu.pk/article/view/61-65.
Section
Original Articles
Author Biographies

ZUJAJA HINA HAROON, CLASSIFIED PATHOLOGIST, COMBINED MILITARY HOSPITAL KOHAT

Lt. COL, ASSISTANT PROFESSOR OF PATHOLOGY

HAROON JAVAID, classified Ophthalmologist,Combined Military Hospital ,Kohat

Assistant Professor,Ophthalmology,Combined Military Hos[pital ,Kohat

ROSS WILLIAMS, Senior Lecturer, Department of Forensic & Biomedical Sciences University of Lincoln, Brayford Pool, Lincoln LN67TS, UK

Senior Lecturer

Department of forensic & Biomedical sciences

Unuversity of Lincoln

YOUSAF KAZMI, Assistant Professor Microbiology Islam Medical College, Sialkot City

Assistant Professor mirobiology

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