Single Stranded Conformation Polymorphism Analysis of the Human p-Defensin 2 Gene in Filipino Tuberculosis Patients

Abstract

Human Beta-Defensin 2 (hBD2) works as a host defense antimicrobial peptide against Mycobacterium tuberculosis, the causative agent of tuberculosis (TB). Genomic DNA from representative samples (TB03 and Cl2) was subjected to polymerase chain reaction (PCR) using the hBD2 Forward and Reverse primers [O.lmM]. Optimized annealing temperature was determined at 69°C in two minutes. Single stranded conformation polymorphism (SSCP) was the technique used to determine the presence of mutations in the hBD2 gene at specific conditions (20% PAGE, 10% APS, and 95% Formaldehye at 4°C). SSCP profiles from ten TB patients and six healthy volunteers demonstrated three specific polymorphism patterns in TB patients and two specific SSCP patterns in the control. These suggest that mutations are present in the hBD2 gene of TB patients.