Aims/Background During the coronavirus disease 2019 (COVID-19) pandemic, the COVID-19 surveillance staff members experienced an increased risk for musculoskeletal disorders. Thus, it is necessary to further investigate the causal factors stemming from COVID-19 surveillance work, especially nucleic acid testing, and establish their relationship with work-related musculoskeletal disorders (WMSDs) by building upon the previous research. This study aimed to determine the prevalence of WMSDs and the major risks faced by the COVID-19 surveillance staff members. Methods This cross-sectional study was conducted in May 2022. The Quick Exposure Check (QEC), Nordic Musculoskeletal Questionnaire (NMQ), and Maslach Burnout Inventory (MBI) were used to evaluate ergonomic factors, musculoskeletal symptoms and pressure among 131 COVID-19 surveillance staff members in four hospitals in South-central Henan Province. Additionally, data regarding the coping strategies adopted by the respondents for lowing the risk of developing WMSDs were collected using a self-report questionnaire. Results The QEC results showed that the back, neck, and should/arm of the COVID-19 surveillance staff members bore high ergonomic loads, while the wrist suffered a medium load. The neck (75.6%), shoulder (66.4%), upper back (58%), and wrist (55.7%) were at higher risk for musculoskeletal disorder. Most of the respondents claimed a low sense of achievement (124, 94.7%) and slightly less than half of them had endured emotional exhaustion at work (61, 46.6%). Prolonged period of maintaining the same posture at work, awkward bending or twisting of back side, task repetition and so on were the perceived job risk factors precipitating WMSDs among the participants in this study. The top three coping strategies for lowering the risk of WMSDs included adjusting the plinth/bed height (49.6%), utilizing different parts of the body at work (47.3%), and taking short breaks when required (33.6%). Conclusion Prolonged large-scale nucleic acid detection can aggravate the musculoskeletal injuries of the neck, shoulder, upper back, and wrist in COVID-19 surveillance staff members. Prevention measures for musculoskeletal injuries factors must be in place to deter such incidence at work.