Detection of HIV-1 RNA in plasma and serum samples using the NASBA amplification system compared to RNA-PCR

J Virol Methods. 1995 Mar;52(1-2):121-32. doi: 10.1016/0166-0934(94)00151-6.


The presence of HIV-1 RNA in the plasma and serum of European and African patients was monitored using RNA-polymerase chain reaction (RNA-PCR) and the new isothermal NASBA nucleic acid amplification system encompassing a gel-based detection assay (ELGA). Identical RNA extraction procedures, provided by the NASBA amplification system, were used for both methods. The detection limit for HIV-1 RNA, measured on a 10-fold dilution series of spiked HIVIIIB in negative plasma, was about 0.05 CCID50 per test for both methods. Both NASBA and RNA-PCR were more sensitive than a p24 assay for the detection of circulating HIV-1 virus in blood: 17 of the 34 (50%) p24 antigen-tested seropositives were p24-positive while 32 (94%) were positive by NASBA and 30 (88%) by RNA-PCR. Among the 45 seropositives, 34 of which were tested for p24 antigen, 43 (96%) were positive by NASBA and 41 (91%) by RNA-PCR. Almost all seropositives had a detectable viral load in 100 microliters plasma. Lower viral loads were only encountered in some healthy seropositives with a higher CD4 count. There was no cross-reactivity with HIV-2 or HIV-I with both the RNA-PCR and NASBA. The extraction method used permitted the detection of HIV-1 RNA equally well in serum and in plasma with heparin or EDTA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Africa / ethnology
  • Algorithms
  • Base Sequence
  • Cell Line
  • Cross Reactions
  • DNA Primers
  • Europe
  • Genes, gag
  • HIV Core Protein p24 / blood
  • HIV Seropositivity / diagnosis*
  • HIV-1 / genetics
  • HIV-1 / isolation & purification*
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques
  • Polymerase Chain Reaction / methods
  • RNA, Viral / blood*
  • Reproducibility of Results
  • Sensitivity and Specificity


  • DNA Primers
  • HIV Core Protein p24
  • RNA, Viral