HIV quantitation in spiked vaginocervical secretions: lack of non-specific inhibitory factors. DATRI 005 Study Team. Division of AIDS Treatment Research Initiative.
The objective of this study was to assess the effect of menstrual phase on the ability to quantitate HIV-1 in vaginocervical secretions (VCS) through reconstruction experiments with HIV seronegative VCS collected throughout the menstrual cycle. Measurement of HIV-1 inoculated into both fresh and frozen VCS was undertaken by quantitative micro co-culture, 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 menstrual cycle and controls); (2) laboratories; (3) stock concentrations; and (4) frozen versus fresh VCS samples. Each assay was assessed independently using a random effects analysis of variance (ANOVA) model. No statistical differences due to menstrual cycle were seen in the assay 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 differences were statistically significant for cell-free RNA (P < 0.001), cell-associated DNA (P < 0.001) and p24 (P < 0.001). It is concluded that VCS obtained throughout the menstrual cycle from HIV-uninfected women lacks intrinsic inhibitory factors which could limit detection and quantification by antigen, culture or nucleic acid-based technologies for HIV-1 in VCS throughout the menstrual cycle. Using a standardized collection procedure, we suggest that variation in HIV quantity over time, when reported in VCS of infected women, should be attributed to HIV-associated biologic factors, rather than non-specific or other technical factors.
Holodniy
M
M
AIDS Research Center, Palo Alto Veterans Affairs Health Care System, CA 94304, USA.
Anderson
D
D
Wright
D
D
Sharma
O
O
Cohn
J
J
Alexander
N
N
Stratton
P
P
Reichelderferd
P
P
D000704:Analysis of Variance; D000956:Antigens, Viral / Q000032:analysis; D001681:Biological Assay; D002582:Cervix Mucus / Q000821:virology*; D015925:Cryopreservation; D005260:Female; D016655:HIV Core Protein p24 / Q000276:immunology; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D008597:Menstrual Cycle; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000302:isolation & purification; D014621:Vagina / Q000378:metabolism / Q000821:virology
D000956:Antigens, Viral; D016655:HIV Core Protein p24; D012367:RNA, Viral