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The development of pathogenic bacteria encompassing antibiotics-resistant genes (ARGs) is a severe environmental concern related to human health. Among all antibiotics administered to humans, the β-lactam group's antibiotics account for two-thirds of the total weight due to its clinical effectiveness and low toxicity. This study investigated the abundance of gene NDM-1 (New Delhi metallo-β-lactamase-1) as an emerging contaminant in Hyderabad, India's water bodies using a quantitative polymerase chain reaction (qPCR). The quantitative microbial risk assessment (QMRA) method was applied to forecast the human health risk linked to ingestion/subjection of surface water with bacteria containing the NDM-1 gene. The gene copy number of the NDM-1 in the exposure route was taken into consideration for the determination of risk in best- and worst-case scenarios. The lowest and highest gene copy numbers were selected as the best-case scenario and the worst-case scenario, respectively. The outlet of Amberpet sewage treatment plant (STP) exhibited the lowest risk, 2×10−2 , i.e., 2 of 100 people are exposed to infection. In contrast, Mir Alam tank lake exhibited the highest risk, 77×10−2 , i.e., 77 of 100 people are exposed to infection. Risk assessment categorized the sampling locations into low, medium, and high risk of infection. Sampling areas with a low risk of infection included Amberpet STP outlet (2×10−2), Ameenpur lake (24×10−3), Manjeera water treatment plant (WTP) outlet (26×10−3), and Osman Sagar lake (24×10−3). Sampling locations with a medium risk of infection included Alwal lake (23×10−2), Durgamcheru lake (47×10−2), Hussain Sagar lake (21×10−2), Mominpet lake (27×10−2), and Saroor Sagar lake (43×10−2). Sampling zones including Fox Sagar lake (53×10−2), Himayat Sagar lake (70×10−2), Kandi lake (56×10−2), Manjeera dam (66×10−2), Mir Alam tank lake (77×10−2), Nagole lake (54×10−2), Safilguda lake (55×10−2), and Singur dam (57×10−2) were categorized as zones with a high risk of infection. The findings of the occurrence of the NDM-1 gene containing bacteria in water bodies surrounding Hyderabad and its related risk will be valuable for developing strategies to safeguard the public from the threat of clinical risks correlated with the dissemination of NDM-1. [ABSTRACT FROM AUTHOR]