A method to detect vulvovaginal candidiasis -associated Candida species and Trichomonas vaginalis in a biological sample, wherein the VVC-associated Candida species comprises Candida glabrata, Candida albicans, Candida tropicalis, C. dubliniensis, C. parapsilosis, Candida krusei, comprising: contacting said biological sample with a plurality of pairs of primers, wherein the plurality of pairs of primers comprises: at least one pair of primers capable of hybridizing to the tefl gene of Candida glabrata, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 20 or SEQ ID NO: 21, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 20 or SEQ ID NO: 21; a plurality of primers capable of hybridizing to the tefl gene of at least one of Candida albicans, Candida tropicalis, C. dubliniensis, and C. parapsilosis, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25; at least one pair of primers capable of hybridizing to the tefl gene of Candida krusei, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 27 or SEQ ID NO: 28, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 27 or SEQ ID NO: 28; and at least one pair of primers capable of hybridizing to the AP-65 gene of Trichomonas vaginalis, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 17 or SEQ ID NO: 18, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 17 or SEQ ID NO: 18; and generating amplicons of the tefl sequences of the Candida species and/or amplicons of the AP-65 gene sequence of Trichomonas vaginalis from said biological sample, if said sample comprises one or more of the VVC-associated Candida species and/or Trichomonas vaginalis; determining the presence or amount of one or more amplified products as an indication of the presence of VVC-associated Candida species and Trichomonas vaginalis in said biological sample. contacting said biological sample with a plurality of pairs of primers, wherein the plurality of pairs of primers comprises: at least one pair of primers capable of hybridizing to the tefl gene of Candida glabrata, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 20 or SEQ ID NO: 21, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 20 or SEQ ID NO: 21; a plurality of primers capable of hybridizing to the tefl gene of at least one of Candida albicans, Candida tropicalis, C. dubliniensis, and C. parapsilosis, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25; at least one pair of primers capable of hybridizing to the tefl gene of Candida krusei, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 27 or SEQ ID NO: 28, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 27 or SEQ ID NO: 28; and at least one pair of primers capable of hybridizing to the AP-65 gene of Trichomonas vaginalis, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 17 or SEQ ID NO: 18, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 17 or SEQ ID NO: 18; and at least one pair of primers capable of hybridizing to the tefl gene of Candida glabrata, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 20 or SEQ ID NO: 21, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 20 or SEQ ID NO: 21; a plurality of primers capable of hybridizing to the tefl gene of at least one of Candida albicans, Candida tropicalis, C. dubliniensis, and C. parapsilosis, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO: 25; at least one pair of primers capable of hybridizing to the tefl gene of Candida krusei, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 27 or SEQ ID NO: 28, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 27 or SEQ ID NO: 28; and at least one pair of primers capable of hybridizing to the AP-65 gene of Trichomonas vaginalis, wherein each primer in said at least one pair of primers comprises a sequence of SEQ ID NO: 17 or SEQ ID NO: 18, or a sequence having one mismatch or two mismatches relative to SEQ ID NO: 17 or SEQ ID NO: 18; and generating amplicons of the tefl sequences of the Candida species and/or amplicons of the AP-65 gene sequence of Trichomonas vaginalis from said biological sample, if said sample comprises one or more of the VVC-associated Candida species and/or Trichomonas vaginalis; determining the presence or amount of one or more amplified products as an indication of the presence of VVC-associated Candida species and Trichomonas vaginalis in said biological sample.