DIAGNOSIS OF HUMAN-IMMUNODEFICIENCY-VIRUS INFECTION USING CITRATED WHOLE-BLOOD

Standard isolation of human immunodeficiency virus type 1 (HIV-1) from peripheral blood mononuclear cells (PBMC) requires 5 to 20 ml of bloo d, and the centrifugal separation of PBMC is expensive and time-consum ing, Whole-blood coculture techniques use small sample volumes, do not require centrifugation, and allow measurement of the total viral burd en in peripheral circulation, We compared the results of citrated whol e-blood coculture with those obtained by the standard AIDS Clinical Tr ials Group PBMC semiquantitative culture method and reverse transcript ion-PCR quantitation of plasma HIV-1 RNA levels, PBMC cocultures were also set up with added erythrocytes (RBCs) to determine if the presenc e of RBCs affects the replication of HIV-1 in vitro, The mean number o f cells required for a p24-positive PBMC coculture was approximately s even times greater than that required for a positive citrated whole-bl ood coculture (P < 0.01), At volumes of 100, 50, and 25 mu l, the sens itivities of the whole-blood coculture were 94.5, 93.6, and 87.3%, res pectively, The PBMC culture in the presence of added RBCs was more sen sitive than PBMC coculture alone, The citrated whole-blood coculture w as simple to perform, produced a reliable diagnosis of HIV infection i n adult volunteers, was more sensitive than previously reported techni ques even in half the culture time, and showed less variability than t he PBMC coculture, Citrated whole-blood coculture may be a useful and efficient tool for diagnosing infection with HIV-1.