Comparative studies on use of fresh and frozen peripheral blood lymphocyte specimens for isolation of human immunodeficiency virus and effects of cell lysis on isolation efficiency.
Heparinized blood specimens (n = 44) and frozen peripheral blood lymphocyte (PBL) specimens (n = 42) were used to evaluate the effects of lysis on human immunodeficiency virus (HIV) isolation. In the two respective groups, 17 and 27 specimens were HIV antibody positive. In the first group there were 8 and in the second group there were 25 that were symptomatic and were classified as indicating an acquired immunodeficiency syndrome-related condition or a pre-acquired immunodeficiency syndrome-related condition by the Centers for Disease Control definition. One-half of the cells from each specimen were frozen and thawed three times before cocultivation with uninfected lymphocytes, and the isolation rates from whole and lysed cells were compared. HIV was isolated from 15 (88%) of 17 fresh specimens and from 24 (89%) of 27 frozen PBLs from HIV antibody-positive patients, and lysis had no overall effect on the isolation rate, which suggested that frozen PBLs were as suitable as fresh blood for HIV isolation attempts and that it was not necessary to maintain cell integrity when submitting PBL samples. Of 21 asymptomatic patients, 20 were culture positive, and of 23 symptomatic patients, 19 were culture positive. Specimens from the 42 antibody-negative individuals were culture negative.
D005615:Freezing; D006678:HIV / Q000201:enzymology / Q000302:isolation & purification*; D006801:Humans; D008214:Lymphocytes / Q000382:microbiology*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism
D012194:RNA-Directed DNA Polymerase