title: Detection of infectious human immunodeficiency virus type 1 in female genital secretions by a short-term culture method.
abstract: Infectious human immunodeficiency virus type 1 (HIV-1) is difficult to detect in female genital secretions by standard virus culture techniques. To improve detection of cell-free HIV-1 in female genital secretions, we adapted a short-term assay that uses the multinuclear-activation galactosidase indicator (MAGI) assay. When vaginal lavages from HIV-1-infected women were tested with the adapted MAGI assay, 25 (64%) of 39 lavages with detectable, cell-free HIV-1 RNA were shown to have infectious virus. No infectious virus was found in 10 vaginal lavages from HIV-1-infected women with undetectable vaginal viral loads. Significantly (P < 0.01) more lavages from HIV-1-infected women tested positive for infectious virus by the MAGI assay than by standard peripheral blood mononuclear cell (PBMC) coculture, which detected infectious virus in only 6 (17%) of 35 vaginal lavages. Lavages with viral loads of >10,000 copies per lavage yielded significantly (P < 0.01) more positive cultures than those with <10,000 copies by using the MAGI assay. Detection of infectious HIV-1 in vaginal lavages was not associated with the presence of genital tract infections or CD4(+)-T-cell counts. However, although the results were not significant (P = 0.08), the MAGI assay detected infectious virus from more vaginal lavages at a vaginal pH of >/=4.5 than at a pH of <4.5. These results indicate that the MAGI assay is more sensitive than PBMC culture methods for detecting infectious virus in female genital secretions. Accurate measurements of infectious virus in genital secretions will improve studies that evaluate sexual transmission of HIV-1.
authors.affiliation: Division of AIDS, STD, and TB Laboratory Research, National Center for Infectious Diseases, National Center for HIV, STD, and TB Prevention, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Cummins
authors.forename: Julie M
authors.initials: JM
authors.lastname: Villanueva
authors.forename: Tammy
authors.initials: T
authors.lastname: Evans-Strickfaden
authors.forename: Shekou M
authors.initials: SM
authors.lastname: Sesay
authors.forename: Sheila R
authors.initials: SR
authors.lastname: Abner
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Bush
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Green
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Lennox
authors.forename: Thomas
authors.initials: T
authors.lastname: Wright
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Folks
authors.forename: Clyde E
authors.initials: CE
authors.lastname: Hart
authors.forename: Charlene S
authors.initials: CS
authors.lastname: Dezzutti
mesh_terms: D000293:Adolescent; D000328:Adult; D018791:CD4 Lymphocyte Count; D002402:Cathepsin D / Q000494:pharmacology; D018920:Coculture Techniques; D003124:Colorimetry; D016207:Cytokines / Q000494:pharmacology; D005260:Female; D005831:Genital Diseases, Female / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008875:Middle Aged; D014621:Vagina / Q000821:virology*; D001616:beta-Galactosidase / Q000032:analysis*
chemical_list: D016207:Cytokines; D001616:beta-Galactosidase; D002402:Cathepsin D