A sensitive microculture assay for the isolation of human immunodeficiency virus type 1 (HIV-1) from blood leukocytes. NLM AIDSLINE Important note: Information in this article was accurate in 1992. The state of the art may have changed since the publication date.

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A sensitive microculture assay for the isolation of human immunodeficiency virus type 1 (HIV-1) from blood leukocytes.

Abstr Annu Meet Am Soc Microbiol. 1991 May 5-9;91:334 (abstract no. T-3). Unique Identifier : AIDSLINE ASM91/0160292
Erice A; Sannerud KJ; Balfour Jr HH; Leske VL; Dept. of Lab Med. & Pathology. Univ. of Minnesota Hosp. and; Minnesota ACTU. Minneapolis, MN.

Abstract: HIV-1 has been isolated by coculturing 10x10(6) peripheral blood mononuclear cells (PBMC) from HIV-1 seropositive patients (pospts) with 5x10(6) PHA-stimulated PBMC from normal donors. Because pospts contain high numbers of HIV-1 infected PBMC, we developed a microculture method (MCX) using fewer PBMC. 1x10(6) PBMC from HIV-1 pospts were cocultivated with 1x10(6) PHA-stimulated donor's PBMC in 1.2 ml of culture medium. Cocultures were incubated for up to 42 days (d). Additional PHA-stimulated donor PBMC (3x10(5)) were added weekly. Culture supernatant was tested every 3-4 d for HIV-1 antigen by EIA (Abbott). 137 MCXs were performed using PBMC from 100 HIV-1 pospts. 139 standard HIV-1 cultures (SCX) from 107 HIV-1 pospts were used as controls. TABULAR DATA, SEE ABSTRACT VOLUME. All +MCX and +SCX were + by d 14. Thus, it is not necessary to maintain HIV-1 cultures for more than 2-3 wks. MCX is a sensitive and less expensive method to recover HIV-1 from PBMC, and is especially important when blood volume is a problem (ie. pediatric pts and AIDS pts with severe anemia).
Keywords: Enzyme-Linked Immunosorbent Assay Human HIV Antigens/ANALYSIS HIV Infections/BLOOD HIV-1/GROWTH & DEVELOPMENT/IMMUNOLOGY/*ISOLATION & PURIF Leukocytes, Mononuclear/DRUG EFFECTS/*MICROBIOLOGY Microbiological Techniques Phytohemagglutinins Virus Cultivation ABSTRACTKWDenzyme-linkedimmunosorbentassayhumanhivantigens/analysishivinfections/bloodhiv-1/growth&development/immunology/KWDisolation&purifleukocytes,mononuclear/drugeffects/KWDmicrobiologymicrobiologicaltechniquesphytohemagglutininsviruscultivationabstract
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