Within the national bovine tuberculosis (bTB) eradication programme in Ireland, concern has been expressed about the quality of testing by veterinarians. However, there is little published evidence supporting this concern, or the challenges that undermine quality testing. Qualitative research methods were used to gather the perspectives of major stakeholders in the bTB eradication (BTE) scheme on the challenges to quality testing for bovine tuberculosis in Ireland. These stakeholders included private veterinarians, government veterinarians, senior managers and herd owners, on the quality of bTB testing and the barriers to improvement. Results are grouped into challenges that exist in the testing environment (ie, at a farm level), and challenges associated with the skills environment (ie, professional skills involved with conducting the test). Challenges in the testing environment include inadequate on-farm testing conditions; lack of clarity on responsibility to ensure adequate testing environment; and the influence of the veterinarian-client relationship. Challenges in the skills environment include deficiencies in the development and supervision of testing skills among trainees and newly qualified veterinarians; and deficiencies in testing standards at a practice level. Regular supervision of testing is necessary to ensure standards. The importance of a continued understanding of the disease (and its eradication) supported by a partnership, cooperative approach between all stakeholders, is emphasised.