Background <p>Needlestick injuries (NSIs) pose a serious occupational risk to healthcare workers. In Pakistan, nurses and nursing students are especially vulnerable, with potential exposure to infections such as HIV and hepatitis B/C. This meta-analysis aimed to estimate the prevalence of NSIs in these groups using data from observational studies conducted across the country.</p> Methods <p>A systematic search was conducted in Medline, EMBASE, CINAHL, Scopus, and Web of Science for studies published between 2000 and 2025. Observational studies reporting NSI prevalence among nurses and nursing students in Pakistan were included. Quality assessment was done using the Joanna Briggs Institute (JBI) tool. A meta-analysis was conducted using a random-effects model due to high heterogeneity. Subgroup analysis, meta-regression, and publication bias assessment were also performed.</p> Results <p>A total of 25 studies with 27 reports were included. The pooled prevalence of NSIs was 48% (95% CI: 35–61%) among nurses and 43% (95% CI: 31–54%) among nursing students. Significant regional differences were found in NSI prevalence among nurses (<i>p</i> = 0.04) and nursing students (<i>p</i> = 0.025) across Sindh, Punjab, and other provinces. Meta-regression showed a significant increase in NSI prevalence among students over time (<i>p</i> = 0.003), with publication bias detected (<i>p</i> = 0.001). The Trim-and-Fill method adjusted the prevalence from 42.8 to 40.4%.</p> Conclusion <p>This meta-analysis provides an estimate of NSI prevalence among nurses and nursing students in Pakistan. The findings highlight the need for effective preventive measures and further research to address the high prevalence of NSIs.</p> Clinical trial number <p>Not applicable.</p>