Most bacteria and fungi are capable of producing biofilms, enabling them to thrive in nature on distinct surfaces. Biofilm formation stands out as one of the most prominent virulence mechanisms that contribute to the infection’s chronicity by functioning as a defense against antimicrobials and host immune systems. Microbial isolates capable of generating biofilms have been discovered to possess higher resistance to frequently administered antifungal drugs. In this research study, 91 Candida isolates from Vulvovaginal Candidiasis (VVC) patients were tested for biofilm development. Candida species were identified, and clinical isolates were tested for antifungal susceptibility (AST). Three methods were used to screen the isolates: the Congo agar method (CRA), the visual tube method (VT), and the Microtitre plate method (MTP). Nearly 60% of the 91 clinical isolates tested were recognized as Non-Albicans Candida (NCAC) species. Itraconazole resistance was shown to be the highest in clinical isolates, followed by Amphotericin B resistance. There were 11(12.09%) isolates that formed strong biofilms, 35(38.46%) isolates that formed moderate biofilms, and 45(49.45%) isolates that formed no biofilm. Because there is a growing incidence of NCAC in the study, it is critical to speciate the Candida species as NCAC are more resistant to routinely used azole medicines. Furthermore, a spike in the prevalence of biofilm producers has been reported, implying greater pathogenicity and antifungal resistance.