title: Quantitative assessment of cell-associated and cell-free virus in cervicovaginal samples of HIV-1-infected women.
abstract: OBJECTIVE
  To examine the amount of cell-free and cell-associated virus in cervicovaginal secretions (CVS) of HIV-infected women.
  
  
  METHODS
  Paired cervicovaginal and blood samples from 61 seropositive women were quantitatively evaluated by competitive polymerase chain reaction (cPCR) and reverse transcription-PCR (cRT-PCR) for: (1) genomic RNA from plasma and cell-free CVS, and (2) unspliced (u/s) RNA transcripts and proviral DNA in cells from secretions.
  
  
  RESULTS
  HIV DNA was detected in 42.6%, u/s transcripts in 32.7% and cell-free HIV RNA in 31.1% of 61 cervicovaginal samples. The median copy numbers of HIV DNA, u/s transcripts, and cell-free RNA were 125 copies/10(5) cells, 40 copies/10(5) cells, and 300 copies/mL of secretion, respectively. Nineteen of 26 (73.1%) and 17 of 26 (65.3%) women positive for DNA were also positive for RNA transcripts and cell-free RNA, respectively (P<0.001). A significant correlation between the amounts of cell-free and u/s transcripts was also found (Spearman Rho 0.618, P=0.014). The prevalences of u/s transcripts and cell-free RNA were 42.6% and 53.8% respectively among patients with detectable blood RNA, and 22.9% (P=0.09) and 14.3% (P=0.0017) among patients with undetectable blood RNA. In stepwise logistic regression, cell-free RNA was independently associated with the presence of detectable blood viremia. The amount of HIV DNA was lower among subiects currently under treatment (50 copies/10(5) cells) than in untreated subjects (250 copies/10(5) cells) (P=0.037).
  
  
  CONCLUSIONS
  Both cell-free and cell-associated HIV could be detected and quantitated in CVS, providing a means to examine the level of viral activity in the female genital tract.
authors.affiliation: Department of SMEC, Section of Microbiology, University of PaviaDepartment of SMEC, Section of Obstetrics and GynecologyDepartment of SMEC, Section of Infectious and Tropical Diseases, University of Pavia and IRCCS S. MatteoDivision of Infectious Diseases, IRCCS S. Matteo, Pavia, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Debiaggi
authors.forename: A
authors.initials: A
authors.lastname: Spinillo
authors.forename: F
authors.initials: F
authors.lastname: Zara
authors.forename: A
authors.initials: A
authors.lastname: Santolo
authors.forename: R
authors.initials: R
authors.lastname: Brerra
authors.forename: R
authors.initials: R
authors.lastname: Maserati
authors.forename: E
authors.initials: E
authors.lastname: Romero
authors.forename: G
authors.initials: G
authors.lastname: Filice