A quality of life questionnaire is rarely adapted to an interview mode for people who mainly use spoken language in daily life. In Taiwan, the WHOQOL-BREF (Mandarin Chinese version) has been developed, as a self-administered questionnaire, but it cannot be applied to the majority of the elderly in Taiwan, who speak only Taiwanese (a dialect). This study adopted the audio player-assisted interview mode to develop a Taiwanese version of the WHOQOL-BREF specifically for Taiwanese-speaking elderly people, and followed with examinations of the reliability and validity of this version. Initially, the WHOQOL-BREF (English version) was translated into colloquial Taiwanese, and field tests confirmed the equivalence and appropriateness of the translation. A total of 228 Taiwanese-speaking elderly people were assessed using the Taiwanese interview version, of which 144 subjects were re-assessed two weeks later. Interviewers assessed each subject aided by an audio player on which all the translated WHO-QOL-BREF contents were recorded. The Taiwanese interview version of the WHOQOL-BREF, except for the item related to dependence on medication, showed acceptable reliability (internal consistency, corrected item-domain correlation, and test-retest reliability) and validity (criterion-related, convergent, and discriminant validity). Confirmatory factor analyses supported the four-factor model of the Taiwanese interview version, providing evidence for construct validity. The results suggest that the Taiwanese audio player-assisted interview version of the WHOQOL-BREF was reliable and valid in assessing quality of life of elderly Taiwanese.