title: Detection and quantitation of human immunodeficiency virus type-1 particles by confocal microscopy.
abstract: A method is described to visualise directly human immunodeficiency virus type-1 (HIV-1) particles. HIV-1 containing samples were adsorbed onto a plastic surface and doubly labeled with antibodies specific for viral proteins and sensitive nucleic acids dyes. Laser scanning confocal microscopy detected co-localization of viral proteins and nucleic acids, thus allowing specific identification of HIV. Using this technique, we have quantified eight different HIV-1 sub-types and three HIV-1 groups in tissue culture supernatants from infected peripheral blood mononuclear cells (PBMCs). Confocal counts correlated well with electron microscopy (EM) counts and HIV-1 RNA loads as determined by quantitative PCR. Confocal microscopy may prove to be a simple alternative to electron microscopy for virus identification and quantitation.
authors.affiliation: Department of Immunology and Molecular Pathology, Windeyer Institute of Medical Sciences, Medical School, Royal Free and University College London, 46 Cleveland Street, London W1T 4JF, UK.
authors.forename: Bishnupriya
authors.initials: B
authors.lastname: Bhattacharya
authors.forename: Robin A
authors.initials: RA
authors.lastname: Weiss
authors.forename: Clare
authors.initials: C
authors.lastname: Davis
authors.forename: Harvey
authors.initials: H
authors.lastname: Holmes
authors.forename: David
authors.initials: D
authors.lastname: Hockley
authors.forename: Ariberto
authors.initials: A
authors.lastname: Fassati
mesh_terms: D002460:Cell Line; D005456:Fluorescent Dyes; D015483:HIV Antibodies / Q000276:immunology; D015497:HIV-1 / Q000145:classification / Q000302:isolation & purification*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology*; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000032:analysis / Q000276:immunology
chemical_list: D005456:Fluorescent Dyes; D015483:HIV Antibodies; D012367:RNA, Viral; D014764:Viral Proteins