Background: Little is known about the engagement of pharmacy assistants (PA) in public health service provision. Objective: To explore the experiences of PA participating in a study to determine whether a cash reward, offered to consumers and pharmacy businesses, increased participation in community pharmacy-based chlamydia screening. Methods: PA experience of the study education and training package, participant recruitment and conducting screening (providing information about chlamydia, specimen collection and handling urine samples) were evaluated using knowledge assessment, a questionnaire and focus groups. Results: Twenty PA participated in the study: 15 (75%) completed all education and training components, 20 (100%) completed the questionnaire and 10 (50%) attended a focus group. PA rated all education and training components as effective (mean visual analog scale scores >8.5). Most PA (13/18, 72.2%) did not support/were unsure about continuing the program, citing the 25% repeat testing rate (presumed to relate to the cash reward) and privacy/confidentiality issues as reasons. Qualitative analysis suggested that minimizing repeat testing, improved workload management and recognition of, and remuneration for, education and training would make this model more acceptable to PA. Conclusion: Findings from this study support the assertion that PA can play a significant role in public health initiatives.