HIV-1 isolation from small amounts of whole blood: a technical evaluation.
We have evaluated a simple and sensitive culture technique for isolation of Human Immunodeficiency Virus Type-1 (HIV-1) from small amounts of whole blood. Data shown in the paper demonstrate that: 1) cell cultures from small amounts of heparinized whole blood (HWB) allow a high isolation rate in infected subjects at all stages of diseases; 2) among asymptomatic subjects the HIV-1 isolation rate is increased in cell cultures from HWB, with respect to cell cultures from peripheral blood mononuclear cells; 3) cultural results from HWB are not influenced by the presence of detectable serum p24 antigen, but a good correlation was found with the titre of anti p24 antibodies in serum; 4) continuous cell lines (such as Molt-3 cells) instead of peripheral blood mononuclear cells can be used, obtaining good results, for HIV-1 isolation from HWB; 5) frozen samples of HWB can be used in cell cultures for HIV-1 isolation; 6) the type of anticoagulant (Heparin or EDTA) used for the collection of blood does not influence viral replication in cell cultures from whole blood; 7) viral isolation from HWB is highly sensitive; amounts so small as five microliters of whole blood are sufficient, in some cases, to obtain viral replication in cell cultures; 8) the minimal dose of HWB sufficient to infect cell cultures (HWB M.D.I.) varied among different patients. Although this work failed to establish a correlation between this parameter and the clinical and immunological status of patients, it is conceivable that HWB M.D.I. could give information about viral load in blood and have a prognostic significance; 9) the HWB M.D.I. rise in patients treated with Zidovudine, suggesting that this method could be employed in the virological evaluation of trials with antiretroviral drugs.
Fiore
J R
JR
Clinic of Infectious Diseases, University of Bari, Italy.
Angarano
G
G
Fico
C
C
Di Stefano
M
M
Grottola
A
A
Monno
L
L
Fracasso
C
C
Pastore
G
G
D000163:Acquired Immunodeficiency Syndrome / Q000188:drug therapy / Q000382:microbiology; D002460:Cell Line; D002478:Cells, Cultured; D015925:Cryopreservation; D004492:Edetic Acid / Q000494:pharmacology; D005069:Evaluation Studies as Topic; D015483:HIV Antibodies / Q000097:blood; D016655:HIV Core Protein p24 / Q000097:blood / Q000276:immunology; D015658:HIV Infections / Q000188:drug therapy / Q000382:microbiology*; D015497:HIV-1 / Q000302:isolation & purification* / Q000502:physiology; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D012680:Sensitivity and Specificity; D013997:Time Factors; D014766:Viremia / Q000382:microbiology*; D014779:Virus Replication; D015215:Zidovudine / Q000627:therapeutic use
D015483:HIV Antibodies; D016655:HIV Core Protein p24; D015215:Zidovudine; D006493:Heparin; D004492:Edetic Acid