Clinical procedures.

Twice per week, the participants attended classes focused on HIV-1 infection prevention, personal empowerment, and job skills training. At each visit, they underwent a finger prick blood draw for quantitative HIV-1 RNA testing. Every 3 months, the participants had a peripheral blood draw and pelvic examination (not performed during menstruation) that included the collection of CVL fluid for sampling from the FGT. CVL fluid was obtained by washing the cervicovaginal walls with 5 ml of sterile saline, which was then centrifuged at 1,700 rpm at 4°C to pellet cells, and the cell-free supernatant, containing free viral particles, was used for RNA sequencing and further analysis in the study (18,–20). The participants also completed a detailed HIV-1 risk questionnaire, which was administered by a counselor; it included STI history, sexual behavior, family planning, use of antibiotics, and diet. Upon detection of a positive HIV-1 RNA test result, the participants underwent blood collection and pelvic examinations with CVL sample collection at 1, 2, 3, 5, 9, 12, 24, 36, and 48 weeks postdetection.