Quantitation and profiling of vaginal microflora

1. A kit comprising:

(a) a first forward primer, a first reverse primer and a first fluorescently-labeled probe against Gardnerella vaginalis , said first forward primer consisting of SEQ ID NO: 7, said first reverse primer consisting of SEQ ID NO: 8, and said first probe consisting of SEQ ID NO: 9;

(b) a second forward primer, a second reverse primer, and a second fluorescently-labeled probe against Lactobacillus iners , said second forward primer consisting of SEQ ID NO: 13, said second reverse primer consisting of SEQ ID NO: 14, and said second probe consisting of SEQ ID NO: 15; and

(c) an instruction for profiling vaginal microflora in a woman comprising instructions for calculating the relative amount of Lactobacillus iners and Gardnerella vaginalis in a biological sample obtained from the woman.

2. The kit of claim 1 , further comprising:

(d) a third forward primer, a third reverse primer and a third fluorescently-labeled probe against Atopobium vaginae , said third forward primer consisting of SEQ ID NO: 1, said third reverse primer consisting of SEQ ID NO: 2, and said third probe consisting of SEQ ID NO: 3;

(e) a fourth forward primer, a fourth reverse primer and a fourth fluorescently-labeled probe against BVAB2, said fourth forward primer consisting of SEQ ID NO: 4, said fourth reverse primer consisting of SEQ ID NO: 5, and said fourth probe consisting of SEQ ID NO: 6;

(f) a fifth, sixth and seventh fluorescently-labeled probes against Lactobacillus crispatus, Lactobacillus gasseri and Lactobacillus jensenii respectively, said fifth probe consisting of SEQ ID NO: 16, said sixth probe consisting of SEQ ID NO: 17 and said seventh probe consisting of SEQ ID NO: 18; and

(g) a fifth forward primer, a fifth reverse primer against Megasphaera , and an eighth and ninth fluorescently-labeled probe against Megasphaera Type 1 and Megasphaera Type 2, respectively, said fifth forward primer consisting of SEQ ID NO: 19, said fifth reverse primer consisting of SEQ ID NO: 20, and said eighth probe consisting of SEQ ID NO: 21 and said ninth probe consisting of SEQ ID NO: 22.

3. A method of diagnosing transitional vaginal microflora status in a woman, comprising the steps of:

(a) quantifying the amount of individual microflora for Lactobacillus crispatus, Lactobacillus gasseria, Lactobacillus jensenni, Lactobacillus iners, Gardnerella vaginalis, Atopobium vaginae, Megasphaera Type 1 , Megasphaera Type 2 and BVAB2 in a biological sample obtained from a woman,

wherein said quantifying step is performed by using a molecular assay selected from the group consisting of qPCR and multiplex qPCR;

(b) calculating the sum of said amount of individual microflora in said biological sample to obtain a total amount of microflora;

(c) calculating the sum of said amount of Lactobacillus iners and Gardnerella vaginalis in said biological sample;

(d) calculating the relative amount of said Lactobacillus iners and Gardnerella vaginalis in said biological sample by dividing said sum amount of Lactobacillus iners and Gardnerella vaginalis by said total amount of microflora,

wherein, said relative amount of Lactobacillus iners and Gardnerella vaginalis , when exceeding 50% of said total amount of microflora, is indicative of a transitional vaginal microflora; and

(e) diagnosing said woman with transitional vaginal microflora status based on said relative amount of Lactobacillus iners and Gardnerella vaginalis in said biological sample;

wherein:

the quantifying of Lactobacillus crispatus is performed using a multiplex qPCR and SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 16;

the quantifying of Lactobacillus gasseria is performed using a multiplex qPCR and SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 17;

the quantifying of Lactobacillus jensenni is performed using a multiplex qPCR and SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 18;

the quantifying of Lactobacillus iners is performed using a multiplex qPCR and SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15;

the quantifying of Atopobium vaginae is performed using SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3;

the quantifying of BVAB2 is performed using SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6;

the quantifying of Gardnerella vaginalis is performed using SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9;

the quantifying of Megasphaera Type 1 is performed using a multiplex qPCR and SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21; and

the quantifying of Megasphaera Type 2 is performed using a multiplex qPCR and SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 22.