Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1993 Aug;31(8):1980–1986. doi: 10.1128/jcm.31.8.1980-1986.1993

Cell-free plasma human immunodeficiency virus type 1 titer assessed by culture and immunocapture-reverse transcription-polymerase chain reaction.

R W Coombs 1, D R Henrard 1, W F Mehaffey 1, J Gibson 1, E Eggert 1, T C Quinn 1, J Phillips 1
PMCID: PMC265683  PMID: 8370724

Abstract

The relationship between plasma human immunodeficiency virus type 1 (HIV-1) infectious titer, determined by quantitative fivefold end-point dilution culture, and the detection of genomic HIV-1 RNA by immunocapture-cDNA-polymerase chain reaction was determined. The optimal plasma specimen collection and storage conditions for the use of such virologic markers for clinical trials were also determined. The variabilities in the measurement of infectious HIVLAI titer associated with intra- and interdonor peripheral blood mononuclear cells were 1.2 and 0.86 log10 50% tissue culture infective doses (TCID50)/ml (95% confidence interval range), respectively. Plasma HIV-1 titers did not change significantly after storing whole blood for 6 h either at 4 degrees C or ambient temperature or plasma for a median of 267 days (range, 259 to 482) at -70 degrees C. The detection of genomic HIV-1 RNA encapsulated in viral particles was very consistent, reproducible, and unaffected by either heparin or acid citrate or by multiple freeze-thawing. The HIV-1 RNA titers also appeared to generally correlate with the biologic titer obtained by the microculture assay. The consistency in infectious HIV-1 titer was evaluated by using 27 unfrozen plasma specimens collected from five subjects over 1 to 9 days. The median change in HIV-1 titer relative to baseline was -0.5 log10 TCID50/ml (interquartile range, -1.03 to 0.175 log10). In contrast, no significant change in HIV-1 RNA for the same frozen plasma specimens was noted. As such, immunocapture-cDNA-polymerase chain reaction may be a useful measure of plasma viremia for studying the natural history of HIV disease and assessing response to therapy.

Full text

PDF
1985

Images in this article

1982Image
on p.1982
1982Image
on p.1982
1983Image
on p.1983
1983Image
on p.1983
1984Image
on p.1984

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albert J., Gaines H., Sönnerborg A., Nyström G., Pehrson P. O., Chiodi F., von Sydow M., Moberg L., Lidman K., Christensson B. Isolation of human immunodeficiency virus (HIV) from plasma during primary HIV infection. J Med Virol. 1987 Sep;23(1):67–73. doi: 10.1002/jmv.1890230108. [DOI] [PubMed] [Google Scholar]
  2. Coombs R. W., Collier A. C., Allain J. P., Nikora B., Leuther M., Gjerset G. F., Corey L. Plasma viremia in human immunodeficiency virus infection. N Engl J Med. 1989 Dec 14;321(24):1626–1631. doi: 10.1056/NEJM198912143212402. [DOI] [PubMed] [Google Scholar]
  3. Ehrnst A., Sönnerborg A., Bergdahl S., Strannegård O. Efficient isolation of HIV from plasma during different stages of HIV infection. J Med Virol. 1988 Sep;26(1):23–32. doi: 10.1002/jmv.1890260105. [DOI] [PubMed] [Google Scholar]
  4. Henrard D. R., Mehaffey W. F., Allain J. P. A sensitive viral capture assay for detection of plasma viremia in HIV-infected individuals. AIDS Res Hum Retroviruses. 1992 Jan;8(1):47–52. doi: 10.1089/aid.1992.8.47. [DOI] [PubMed] [Google Scholar]
  5. Ho D. D., Moudgil T., Alam M. Quantitation of human immunodeficiency virus type 1 in the blood of infected persons. N Engl J Med. 1989 Dec 14;321(24):1621–1625. doi: 10.1056/NEJM198912143212401. [DOI] [PubMed] [Google Scholar]
  6. Holodniy M., Katzenstein D. A., Sengupta S., Wang A. M., Casipit C., Schwartz D. H., Konrad M., Groves E., Merigan T. C. Detection and quantification of human immunodeficiency virus RNA in patient serum by use of the polymerase chain reaction. J Infect Dis. 1991 Apr;163(4):862–866. doi: 10.1093/infdis/163.4.862. [DOI] [PubMed] [Google Scholar]
  7. Holodniy M., Kim S., Katzenstein D., Konrad M., Groves E., Merigan T. C. Inhibition of human immunodeficiency virus gene amplification by heparin. J Clin Microbiol. 1991 Apr;29(4):676–679. doi: 10.1128/jcm.29.4.676-679.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Katzenstein D. A., Holodniy M., Israelski D. M., Sengupta S., Mole L. A., Bubp J. L., Merigan T. C. Plasma viremia in human immunodeficiency virus infection: relationship to stage of disease and antiviral treatment. J Acquir Immune Defic Syndr. 1992;5(2):107–112. [PubMed] [Google Scholar]
  9. Li Y., Kappes J. C., Conway J. A., Price R. W., Shaw G. M., Hahn B. H. Molecular characterization of human immunodeficiency virus type 1 cloned directly from uncultured human brain tissue: identification of replication-competent and -defective viral genomes. J Virol. 1991 Aug;65(8):3973–3985. doi: 10.1128/jvi.65.8.3973-3985.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Michael N. L., Vahey M., Burke D. S., Redfield R. R. Viral DNA and mRNA expression correlate with the stage of human immunodeficiency virus (HIV) type 1 infection in humans: evidence for viral replication in all stages of HIV disease. J Virol. 1992 Jan;66(1):310–316. doi: 10.1128/jvi.66.1.310-316.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Moore J. P., McKeating J. A., Weiss R. A., Sattentau Q. J. Dissociation of gp120 from HIV-1 virions induced by soluble CD4. Science. 1990 Nov 23;250(4984):1139–1142. doi: 10.1126/science.2251501. [DOI] [PubMed] [Google Scholar]
  12. Moudgil T., Daar E. S. Infectious decay of human immunodeficiency virus type 1 in plasma. J Infect Dis. 1993 Jan;167(1):210–212. doi: 10.1093/infdis/167.1.210. [DOI] [PubMed] [Google Scholar]
  13. Piatak M., Jr, Saag M. S., Yang L. C., Clark S. J., Kappes J. C., Luk K. C., Hahn B. H., Shaw G. M., Lifson J. D. High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR. Science. 1993 Mar 19;259(5102):1749–1754. doi: 10.1126/science.8096089. [DOI] [PubMed] [Google Scholar]
  14. Resnick L., Veren K., Salahuddin S. Z., Tondreau S., Markham P. D. Stability and inactivation of HTLV-III/LAV under clinical and laboratory environments. JAMA. 1986 Apr 11;255(14):1887–1891. [PubMed] [Google Scholar]
  15. Saag M. S., Crain M. J., Decker W. D., Campbell-Hill S., Robinson S., Brown W. E., Leuther M., Whitley R. J., Hahn B. H., Shaw G. M. High-level viremia in adults and children infected with human immunodeficiency virus: relation to disease stage and CD4+ lymphocyte levels. J Infect Dis. 1991 Jul;164(1):72–80. doi: 10.1093/infdis/164.1.72. [DOI] [PubMed] [Google Scholar]
  16. Wang J. T., Wang T. H., Sheu J. C., Lin S. M., Lin J. T., Chen D. S. Effects of anticoagulants and storage of blood samples on efficacy of the polymerase chain reaction assay for hepatitis C virus. J Clin Microbiol. 1992 Mar;30(3):750–753. doi: 10.1128/jcm.30.3.750-753.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES