HIV-1 RNA Quantification with Reflex to HIV-1 Genotypic Drug Resistance Mutation Analysis, Plasma
Method Description Describes how the test is performed and provides a method-specific reference
For Detection and Quantification:
COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, version 2.0, is an in vitro nucleic acid amplification test for the quantification of HIV-1 RNA in human plasma, using the COBAS AmpliPrep instrument for automated viral nucleic acid extraction (generic silica-based capture technique) and the COBAS TaqMan analyzer for automated amplification and detection of the viral nucleic acid target. To accommodate the polymorphism within the HIV-1 genomic target sequence, this assay employs multiple PCR primers for 2 target sequences (HIV-1 gag and LTR regions). The HIV-1 Quantitation Standards is used to detect and compensate for possible PCR inhibition and as a control for target amplification and detection processes. Dual-labeled fluorescent oligonucleotide probes are utilized to detect amplified material. Quantification of target sequences is monitored using the emission intensity of fluorescent reporter dyes released during the amplification process.(Package insert: COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, version 2.0, Roche Molecular Systems Inc., Branchburg, NJ, 6/2010)
For Genotypic Resistance Analysis:
HIV-1 genotyping is performed using the TruGene HIV-1 assay, which determines the nucleotide bases by simultaneous bidirectional sequencing (1.9 kb total) of the viral reverse transcriptase (1,600 nucleotide) and protease (300 nucleotide) genes of HIV-1 in a blood specimen and identifies any mutations. An automated DNA sequencing system (OpenGene computer software) compares the specimen genotype to the known resistance mutations and generates a list of the mutations present and the antiviral drugs to which the mutations confer resistance.(Package insert: TruGene HIV-1 Genotyping Assay, Siemens Healthcare Diagnostics, Tarrytown, NY)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
HIV-1 quantification: Monday through Saturday; 7 a.m.-4 p.m.
HIV-1 genotyping: varies; test will be performed in batches of 4.
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test