HIV QUANTITATION IN SPIKED VAGINOCERVICAL SECRETIONS - LACK OF NONSPECIFIC INHIBITORY FACTORS

The objective of this study was to assess the effect of menstrual phas e on the ability to quantitate HIV-1 in vaginocervical secretions (VCS ) through reconstruction experiments with HIV seronegative VCS collect ed throughout the menstrual cycle. Measurement of HIV-1 inoculated int o both fresh and frozen VCS;was undertaken by quantitative micro co-cu lture, p24 antigen assay and polymerase chain reaction (PCR) for both HIV-1 RNA and pro-viral DNA. Two laboratories carried out these assays over a range of viral concentrations; The study involved a randomized factorial design and the factors were: (1) diluents (phases of the me nstrual cycle and controls); (2) laboratories: (3) stock concentration s; and (4) frozen versus fresh VCS samples. Each assay was assessed in dependently using a random effects analysis of variance (ANOVA) model. No statistical differences due to menstrual cycle were seen in the as say results of p24 antigen (P = 0.08), PBMC culture(P = 0.74), plasma culture (P = 0.13), cell-free RNA (P = 0.44), cell-associated RNA (P = 0.58) and cell-associated DNA (P = 0.43). inter-laboratory difference s were statistically significant for cell-free RNA (P < 0.001), cell-a ssociated DNA (P < 0.001) and p24 (P < 0,001). It is concluded that VC S obtained throughout the menstrual cycle from HIV-uninfected women la cks intrinsic inhibitory factors which could limit detection and quant ification by antigen, culture or nucleic acid-based technologies for H IV-1 in VCS throughout the menstrual cycle. Using a standardized colle ction procedure, we suggest that variation in HIV quantity over time, when reported in VCS of infected women, should be attributed to HIV-as sociated biologic factors, rather than non-specific or other technical factors. (C) 1998 Elsevier Science B.V. All rights reserved.