Cholera epidemic and the occurrence of drug resistant Vibrio cholerae are being reported with increasing frequency world-wide. Pathogenic bacteria develop antibiotic resistance due to the indiscriminate use of antibiotics, especially in developing countries. Changes in the drug susceptibility pattern and genes within SXT-Constin were observed in V.cholerae O1. The spread of resistant strains has been attributed in part of SXT-C. This study is aimed to investigate the regional variation in antibiotic resistance pattern as well as to find out the presence of antibiotic resistance genes, class I integron (intSXT) and SXT-C within V.cholerae O1 isolates which would help us to control the emergence of antimicrobial drug resistance. Antibiotic susceptibility pattern of the clinical isolates of V. cholerae O1 from Bangladesh, Pakistan, Nepal and Zimbabwe were investigated following standard procedures. In this study a total of 60 V.cholerae O1 strains (15 from each country) were subjected to antibiotic susceptibility testing against 12 commonly used antibiotics. For further molecular analysis a total of 40 strains (10 from each country) were studied for the presence of antibiotic resistance genes, Class I integron and SXT-C. All of the isolates which were used in this study were collected from cholera pandemic Bangladesh (during cyclone Aila, 2009); cholera epidemic Pakistan (2010), Nepal (2009) and Zimbabwe (2009). The antibiogram results revealed that all the isolates from Bangladesh (BD), Pakistan (PK), Nepal (N) and Zimbabwe (Z) showed resistance towards Sulfamethoxazole-Trimethoprime (SXT). Isolates from BD showed 100% resistance towards Polymixin B (PB), Naldixic acid (NA) and Streptomycin (S); while isolates from N, PK and Z showed 100% resistance towards S, NA & S respectively. About 40%, 33.33% and 53.3% isolates of PK, N and Z respectively showed resistance pattern towards PB. Isolates from N and Z respectively showed 100% & 86.6% resistance towards NA; while 86.6% isolates of PK showed resistance towards S. Isolates from BD (100%) showed resistance towards Chloramphenicol (C) while 13.3%, 73.3% & 66.6% of isolates from PK, N and Z respectively showed resistance towards C. Isolates form BD, PK, N and Z showed 80%, 44.4%, 54.4% and 100% resistance pattern respectively towards Tetracycline (TE). All of the isolates from different countries were sensitive towards Azythromycin (AZM), Ceftriaxone (CRO), Cefepime (FEP), Gentamycin (CN) and Ciprofloxacin (CIP) except one isolate each from BD & PK was intermediate resistant towards CRO & FEP respectively and other four isolates from PK were resistant towards AZM. The highest Multiple Antibiotic Resistance (MAR) index was observed 0.58 for isolates from Pakistan and Nepal and lowest was 0.16 observed for the isolates of Pakistan, Nepal and Zimbabwe. Among 40 isolates 27 (67.5%) were positive for the presence of Class I integron (intSXT). However, none of the integron-positive isolates belonged to the group of SXT or R391. Isolates from Nepal were positive for the presence of sulII, strA and floR, while isolates from Bangladesh, Pakistan and Zimbabwe were only positive for sulII, strA; sulII, floR and strA, floR respectively. All the isolates from 4 countries were positive for the presence of traI, traC,setR and traG-s07. Cds38-traL was found within isolates of Bangladesh, Pakistan and Zimbabwe while s075-traF and kan claster were only