Background and Purpose: Opportunistic fungal infections have been on a growingtrend since the last two decades. Among the opportunistic fungal agents, Candidaspecies, Cryptococcus neoformans, and Aspergillus fumigatus account for most of thelife-threatening infections in immunocompromised individuals. Regarding this, thepresent study aimed to investigate the molecular identification and antifungalsusceptibility pattern of Candida species isolated from HIV-infected patients.Materials and Methods: This study was conducted on 80 clinical samples collectedfrom HIV-infected patients with suspected candidiasis referring to Tagore MedicalCollege and Hospital, Rathinamangalam and Government Hospital of ThoracicMedicine, in Chennai, India, for 18 months (i.e., May 2016-December 2017). Phenotypicand molecular identification was accomplished using internal transcribed spacer region 1(ITS1) and ITS4 primers. The antifungal susceptibility pattern of the isolates against fourantifungal agents was also determined by both disk diffusion and broth dilution methods.Results: In the present study, the prevalence of candidiasis was obtained as 75% (n=60).Candida tropicalis was the predominant identified species. All the emerging species(i.e., Kodamaea ohmeri, Hanseniaspora opuntiae, and C. orthopsilosis) were identifiedthrough molecular identification since the phenotypic identification was inconclusive. Interms of the susceptibility pattern, 63.3% and 18.3% of the isolates were resistant tofluconazole and voriconazole, respectively. Candida albicans was also found to beresistant to amphotericin B.Conclusion: Molecular assay led to the identification of K. ohmeri, H. opuntiae, and C.orthopsilosis, which were multidrug-resistant. This study highlighted the need for theprompt and timely identification of clinical yeast isolates given the emergence of manyrare species and their capability of causing life-threatening infections and outbreaks. Inthe laboratories where molecular diagnostic methods are not available, alternativeservices of reference laboratories can be utilized as cost-effective measures. With regardto the growing prevalence of antifungal drug resistance, antifungal susceptibility testingshould be made mandatory for effective patient management.