title: Diagnosis of human immunodeficiency virus infection using citrated whole blood.
abstract: Standard isolation of human immunodeficiency virus type 1 (HIV-1) from peripheral blood mononuclear cells (PBMC) requires 5 to 20 ml of blood, and the centrifugal separation of PBMC is expensive and time-consuming. Whole-blood coculture techniques use small sample volumes, do not require centrifugation, and allow measurement of the total viral burden in peripheral circulation. We compared the results of citrated whole-blood coculture with those obtained by the standard AIDS Clinical Trials Group PBMC semiquantitative culture method and reverse transcription-PCR quantitation of plasma HIV-1 RNA levels. PBMC cocultures were also set up with added erythrocytes (RBCs) to determine if the presence of RBCs affects the replication of HIV-1 in vitro. The mean number of cells required for a p24-positive PBMC coculture was approximately seven times greater than that required for a positive citrated whole-blood coculture (P < 0.01). At volumes of 100, 50, and 25 microl, the sensitivities of the whole-blood coculture were 94.5, 93.6, and 87.3%, respectively. The PBMC culture in the presence of added RBCs was more sensitive than PBMC coculture alone. The citrated whole-blood coculture was simple to perform, produced a reliable diagnosis of HIV infection in adult volunteers, was more sensitive than previously reported techniques even in half the culture time, and showed less variability than the PBMC coculture. Citrated whole-blood coculture may be a useful and efficient tool for diagnosing infection with HIV-1.
authors.affiliation: Department of Molecular Microbiology and Immunology, Johns Hopkins University School of Public Health, Baltimore, Maryland 21205, USA.
authors.forename: U K
authors.initials: UK
authors.lastname: Sharma
authors.forename: H F
authors.initials: HF
authors.lastname: Song
authors.forename: F F
authors.initials: FF
authors.lastname: Willingham
authors.forename: J
authors.initials: J
authors.lastname: Hannig
authors.forename: C
authors.initials: C
authors.lastname: Flexner
authors.forename: H
authors.initials: H
authors.lastname: Farzadegan
authors.forename: C
authors.initials: C
authors.lastname: Nicolau
authors.forename: D H
authors.initials: DH
authors.lastname: Schwartz
mesh_terms: D000328:Adult; D019343:Citric Acid; D005069:Evaluation Studies as Topic; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods* / Q000706:statistics & numerical data
chemical_list: D012367:RNA, Viral; D019343:Citric Acid