To address a severe shortage of human resources for health, the Zambian Ministry of Health has begun to make use of lay counsellors for HIV counselling and testing. However, their skills and knowledge rarely have been reviewed or refreshed. We conducted a two-day refresher workshop for lay counsellors to review their performance and refresh their skills and knowledge. The objective of this study was to evaluate the refresher training intervention for HIV lay counsellors in the rural district of Chongwe in Zambia. The two-day refreshertraining workshop was held in November 2009. Twenty-five lay counsellors were selected by District Health Office and participated in the workshop. The workshop included: the opening, a pre-training exercise, lectures on quality assurance with regard to testing and safety precautions, lectures on counselling, filling the gap/Q&A session, and a post-training exercise. In both the pre- and post-trainingexercise, participants answered 25 true/false questions and tested 10 blood panel samples to demonstrate their knowledge and skill on HIV counselling and testing. The average overall knowledge test score  increased from 79% to 95% (p<0.001). At the baseline, knowledgetest scores in topic of standard precaution and post-exposure prophylaxis were relatively low (58%) but rose to 95% after the training (p<0.001). The per cent agreement of HIV testing by lay counsellors with reference laboratory was 99.2%. Participants’ knowledge was improved during the workshop and skill at HIV testing was found to remain at a high level of accuracy. Relatively weak knowledge of standard precautions and post-exposure prophylaxis suggests that lay counsellors are at risk of nosocomial infections, particularly in the absence of refresher training interventions. We conclude that the refresher training was effective for improving the knowledge and skills of lay counsellors and provided an opportunity to monitor their performance