Performance characteristics for the quantification of plasma HIV-1 RNA using branched DNA signal amplification technology

Article Abstract:

The bDNA-1.0 test may be preferable for evaluating blood samples for human immunodeficiency virus-1 RNA (HIV-1 RNA) levels compared to the reverse transcription polymerase chain reaction (RT-PCR) test. HIV-1 RNA levels may be useful indicators of disease progression in patients with HIV. Researchers evaluated the accuracy and stability of the bDNA-1.0 test as compared to the RT-PCR test. The bDNA-1.0 test was consistently more accurate and could detect smaller changes in sample variability as compared to the RT-PCR test. The presence of fatty acids, bile pigment, and drugs used to treat HIV had no significant effect on the accuracy of the bDNA-1.0 test. Plasma samples low in hemoglobin and stored in the anticoagulant EDTA seem to generate the best test results. Samples were stable after withstanding extended storage in a -80 degree C freezer and also after subjection to three freezing and thawing cycles.

Usage, Innovations, HIV infection, HIV infections, Genetic aspects, Gene amplification, Immunoassay, Viral genetics

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Rapid and precise quantification of HIV-1 RNA in plasma using a branched DNA signal amplification assay

Article Abstract:

The branched DNA (bDNA) signal amplification assay may be useful in measuring blood levels of HIV RNA during HIV infection. This assay uses nucleotide probes attached to a microwell that bind to viral RNA in blood samples placed in the well. Researchers used the assay to measure HIV RNA in blood samples from 349 HIV-positive people. The assay detected HIV RNA in 83% of the patients with CD4 T cell counts less than 500. HIV RNA levels rose as the number of CD4 T cells dropped. However, RNA levels remained stable in patients with stable disease. HIV RNA levels initially dropped in one patient on long-term AZT therapy, but began to rise again 19 weeks after treatment began. This indicates that the drug was no longer effective, even though the patient's CD4 counts remained stable. This assay could be useful in documenting a patient's response to treatment.

RNA, Virus replication

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Comparison of selective polymerase chain reaction primers and differential probe hybridization of polymerase chain reaction products for determination of relative amounts of codon 215 mutant and wild-type HIV-1 populations

Article Abstract:

A technique known as differential hybridization may offer advantages over selective polymerase chain reaction (PCR) in detecting protein mutations associated with increases in the amount of HIV in the body. Researchers sought to identify protein mutations in HIV-infected patients receiving zidovudine to see if the drug was working. The first technique, selective PCR, analyzed protein mutations in the RNA of HIV from blood plasma. Selective PCR was found to be 80% sensitive in detecting mutations but technical difficulties hindered the experiment. A second test, differential hybridization of PCR product with radiolabelled probes, detected mutations in 100% of 127 blood samples. Increases in protein mutation correlated with increases in the amount of HIV.

Mutation (Biology), Mutation

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