The present study deals with the determination of benzo(a)pyrene in biological samples and detection of genetic damage in the subjects working at petrol pumps. The multidisciplinary study involved questionnaire survey and analysis of biological samples. The questionnaire survey was carried out in 107 petrol pumps of Lahore and Gujrat districts for the assessment of health status of the workers (n=1000) and risk factors in their workplace settings. The biochemical tests included; detection of benzo(a)pyrene in blood and hair samples by HPLC (n=200). Buccal micronucleus cytome assay (BMCyt) was carried out for the identification of nuclear anomalies i.e. micronucleus (MN), binucleus (BN) for the evaluation of genotoxic parameters and cytotoxic damage by karyorrhexis (KR) and karyolysis (KL) cells. The DNA damage was further assessed by comet assay through total comet scoring (TCS). Genetic polymorphism was detected in associated genes in selected blood samples of the petrol pump workers through polymerase chain reaction (PCR). Data was analyzed through ANOVA, t-test (p<0.05) and Pearson correlation. Questionnaire survey revealed the study respondents had low socioeconomic and educational status, poor dietary habits, various health problems and specially no use of PPE during work. Analysis of samples showed significantly higher concentration of BaP in blood (µg/l) and hair (µg/g), frequencies of buccal cell anomalies and TCS in the petrol pump workers (exposed) as compared to non-exposed subjects. The PCR of the selected samples evidently revealed polymorphism in the associated genes (CYP1A1 and GSTM1) hence pointing towards the more probability of susceptibility to diseases related to lungs. The results were compared on the basis of age, job duration, smoking habits, working hours per day and job types. The subjects of higher ages, long job duration, smokers and longer working hours per day were found with significantly higher BaP concentration in blood and hair. Similarly the buccal cell anomalies and total comet scores was also significantly higher in these subjects. The BaP concentration in blood was strongly corelated to the MN frequencies in buccal cells and TCS in blood cells of the petrol pump workers. The results are depictive of the fact that low socioeconomic status, less education, lack of awareness, no use of PPE, personal hygiene and smoking habits appear to be the contributory factors. Results of BaP in blood and hair revealed that the petrol pump workers are likely to get occupational exposure to PAHs like BaP and other such substances in their workplace because of many reasons especially due to ignorance, nonuse of PPE, in effective regulations, monitoring as well as the faulty working practices. The results of the study are significant and provide enough evidence with reference to the health and safety issues at the petrol pumps in Pakistan. On the basis of the results; recommendations are drawn for the future work with reference to health risk assessment and management at work. It also advocates for the formulation of specific rules for petrol pump workers.