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Dhat syndrome is a culture-bound syndrome, characterized by the core belief of loss of semen accompanied by symptoms of general weakness, lack of energy and concentration, impaired sexual functions, and vague somatic troubles, often associated with an anxious or dysphoric mood state. Although many studies have described the clinical picture of Dhat syndrome, there is lack of availability of an instrument which can comprehensively assess patients presenting with Dhat syndrome.The aim of this article is to develop a questionnaire that can comprehensively assess Dhat syndrome and guide the clinicians in managing such patients.Initially, an extensive literature review was done to prepare a comprehensive list of clinical features, beliefs associated with the passage of “Dhat,” and attribution of the symptoms by the patients. These items were converted into a questionnaire for investigational interview. The questionnaire thus developed was administered to 54 patients and was also sent to eight subject experts for their opinion on the questionnaire.To develop and evaluate the content validity of the Comprehensive Questionnaire for Assessment of Dhat Syndrome.As per the opinion of experts, the questionnaire had good content validity and was useful for not only clinicians dealing with patients of Dhat syndrome but was also considered useful for the patients presenting with Dhat syndrome. All the experts found the questionnaire to be comprehensive, and two-third of the experts regarded the length to be adequate. Although none of them suggested any deletion of items, yet some additions were suggested. The language of the questionnaire was rated from simple to very simple. Results of administration of the questionnaire on 54 patients of Dhat syndrome established that the questionnaire was helpful in providing comprehensive clinical picture of Dhat syndrome.The questionnaire designed for the purposes of this study is a useful instrument for comprehensive assessment of the clinical picture of Dhat syndrome. Grover S, Avasthi A, Aneja J, Shankar G, Mohan M R, Nehra R, and Padhy SK. Comprehensive questionnaire for assessment of Dhat syndrome: Development and use in patient population. J Sex Med 2014;11:2485–2495.