Objectives The Karnofsky score (KS) modified for cats, a scoring system to rate health and quality of life (QOL) in cats, is used in clinical trials, but its reliability and validity are yet to be determined. The present study aims to evaluate the scientific robustness of the KS when adapted for use in a hospital setting. Methods A list of variables to consider during the physical examination, which informs the clinician's score (CS) part of the KS, was added and clinicians were allowed to choose a score anywhere between 0 and 50. The Karnofsky QOL questionnaire was adapted for use in a hospital setting. F-tests with Bonferroni correction and Spearman rank correlation coefficients were used to evaluate reliability and validity of the KS to assess the health and wellbeing of cats in a hospital setting. The records of 54 feline immunodeficiency virus-positive cats, which were recruited for a clinical trial and hospitalised for 6 weeks, were reviewed. Four veterinarians scored the CS, and one veterinarian and a veterinary nurse assessed the QOL score. Results Mean absolute difference between observers was significantly larger for the CS than for the QOL score ( P <0.001) and two veterinarians scored significantly higher than the remaining two veterinarians ( P <0.001). Inter-observer correlation ranged from 0.45-0.75 for the CS. For the QOL score, the absolute difference between observers was small, no significant difference was found between observers and a high degree of inter-observer correlation was noted (r = 0.91). Conclusions and relevance The results indicate low inter-observer reliability for the CS, requiring additional modifications to this part of the KS. The QOL score seems more reliable, and the questionnaire may serve as a reliable tool in the assessment of QOL in cats in a hospital setting. Consequently, further adaptation of the KS is mandatory when simultaneous assessment of both the cat's clinical health and perceived wellbeing is required.