Direct quantification of HIV-1 infectivity for monocyte--macrophages using an infectious focus assay.
Monocyte--macrophages are important target cells and reservoirs for HIV. The existing methods for the quantification of infectious virus in HIV stocks are not totally satisfactory for use with macrophage cultures. We have developed an infectious focus assay for the direct quantification of virions infectious for human peripheral blood monocyte-derived macrophages adhering to plastic microtitre plates. The combination of an HIV-1 p24-antigen-specific monoclonal antibody and a beta-galactosidase-linked second antibody resulted in a sensitive and very specific assay. With 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside as substrate, the assay proved to be as sensitive as p24 antigen quantification in culture supernatants.
Sonza
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Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Melbourne Australia.
Burgess
S H
SH
Crowe
S M
SM
D015683:Gene Products, gag / Q000032:analysis; D015488:HIV Antigens / Q000032:analysis; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000254:growth & development* / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D008264:Macrophages / Q000382:microbiology*; D009000:Monocytes / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014758:Viral Core Proteins / Q000032:analysis; D001616:beta-Galactosidase / Q000378:metabolism
D015683:Gene Products, gag; D015488:HIV Antigens; D016655:HIV Core Protein p24; D014758:Viral Core Proteins; D001616:beta-Galactosidase