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Background: In recent years, neonatal hearing screening (NHS) has gained rapid traction in both developed and developing nations. However, the efficacy of these efforts depends on comprehensive standardization across all screening facets. This study aimed to assess the status and quality of NHS by investigating the knowledge, attitudes, beliefs, and practices of hearing screening practitioners regarding NHS. Methods: A cross-sectional survey was conducted, and an online questionnaire based on the knowledge-attitude/belief (A/B)-practice model was distributed to all NHS practitioners in Luzhou, western China. Valid questionnaires were examined and uniformly graded. Results: A total of 63 valid questionnaires were collected. The practitioners were mainly female (96.83%), with nursing backgrounds (63.49%), and undergraduate degrees (66.67%). Most had ≤5 years of experience (74.60%) and had junior/intermediate titles (93.65%). The NHS within the Luzhou area started in 2006 with provincial institutions, expanding to 42 institutions by 2022. Statistically significant correlations were observed between the A/B score and the conducting years of each NHS institution (p <.05) as well as between the Knowledge (K) and Practice (P) scores (p <.01). No significant correlation was found between the K score, P score, A/B score, and working years of practitioners (p >.05), or in the total score of NHS institutions at different levels or in different counties by one-way ANOVA (p >.05). Conclusions: It has been 17 years since the first medical institution in Luzhou launched NHS, and the overall performance of practitioners from different institutions has been consistent in terms of their knowledge, attitudes, or level of practice. However, there is room for further improvement in both the professional development of individuals and aspects related to work, such as health education and long-term follow-up. [ABSTRACT FROM AUTHOR]