Detection and quantitation of human immunodeficiency virus type-1 particles by confocal microscopy.
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.
Bhattacharya
Bishnupriya
B
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.
Weiss
Robin A
RA
Davis
Clare
C
Holmes
Harvey
H
Hockley
David
D
Fassati
Ariberto
A
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
D005456:Fluorescent Dyes; D015483:HIV Antibodies; D012367:RNA, Viral; D014764:Viral Proteins