RealTime HIV-1
- Indications and Limitations of Use
- Package Insert For Information Only - Not a Controlled Copy
The Abbott RealTime HIV-1 assay is an in vitro reverse transcription-polymerase chain reaction (RT-PCR) assay for the quantitation of Human Immunodeficiency Virus type 1 (HIV-1) on the automated m2000 System in human plasma from HIV-1 infected individuals over the range of 40 to 10,000,000 copies/mL. The Abbott RealTime HIV-1 assay is intended for use in conjunction with clinical presentation and other laboratory markers for disease prognosis and for use as an aid in assessing viral response to antiretroviral treatment as measured by changes in plasma HIV-1 RNA levels. This assay is not intended to be used as a donor screening test for HIV-1 or as a diagnostic test to confirm the presence of HIV-1 infection.
| Abbott RealTime HIV-1 Assay | |
| Sensitivity | 40 copies/mL for 1.0 mL sample volume 40 copies/mL for 0.6 mL sample volume 75 copies/mL for 0.5 mL sample volume 150 copies/mL for 0.2 mL sample volume |
| Linear Range | 40 copies/mL (1.6 log copies/mL) to 10 million copies/mL (7.0 log copies/mL) |
| Specificity | 100%1 |
| Target region | Integrase region of polymerase gene |
| Subtype Detection | Group M subtypes A—H, Group O and Group N |
| Internal control | Non-competitive pumpkin RNA, added to lysis buffer during extraction |
| Standardization | Virology Quality Assurance (VQA) Laboratory of the AIDS Clinical Trial Group World Health Organization (WHO) 1st International Standard for HIV-1 RNA (97/656) |
| Specimen Type | Plasma (ACD-A and EDTA) |
| Input volume | 0.2mL; 0.5mL; 0.6mL; 1.0mL |
| Sample preparation | m2000sp |
1 The specificity of the RealTime HIV-1 assay was evaluated at three external sites by testing 514 HIV-1 seronegative plasma specimens from volunteer blood donors. HIV-1 RNA was not detected for all 514 specimens and the RealTime HIV-1 assay specificity was estimated to be 100% (514/514), (95% CI 99.28 to 100%).
| Detection of HIV-1 Subtypes and Groups | ||||
| Group/ Subtypes |
n | RealTime Detected |
Comparator 1 Detected |
Comparator 2 Detected |
| M/Subtype A | 10 | 10 | 10 (1) | 10 (1) |
| M/Subtype B |
10 | 10 | 10 (0) | 10 (0) |
| M/Subtype C |
10 | 10 | 10 (0) | 10 (0) |
| M/Subtype D |
10 | 10 | 10 (0) | 10 (0) |
| M/Subtype AE | 10 | 10 | 10 (0) | 10 (0) |
| M/Subtype F | 10 | 10 | 10 (0) | 10 (0) |
| M/Subtype AG | 10 | 10 | 10 (3) | 10 (1) |
| M/Subtype G | 10 | 10 | 10 (2) | 10 (1) |
| Group O | 10 | 10 | 0 (NA) | 7 (7) |
A total of 90 clinical specimens, ten of each Group M subtype (A, B, C, D, CRF01-AE, F, CRF02-AG, G) and of Group O, were tested with the RealTime HIV-1 assay and by two other approved HIV-1 quantitative assays referred to as Comparator 1 (FDA-approved version used) and Comparator 2 (CE-Marked version used). The numbers in parentheses are the number of specimens that had lower quantitation values by more than 1.00 log copies/mL when compared to RealTime HIV-1 assay.
| Correlation to Comparator Assay |
A total of 301 specimens collected from HIV-1 infected patients were tested with the RealTime HIV-1 assay at three external sites and with the comparator method at a central laboratory site. The results from a total of 259 specimens that fell within the common assay dynamic range were analyzed by the Passing-Bablok linear regression method.
Development Philosophy
Today's clinical molecular diagnostics laboratory must have confidence in the quality of the HIV-1 patient results. As a result of our real-time PCR development philosophy, Abbott RealTime HIV-1 is engineered to tolerate the genetic diversity of HIV-1 Subtypes.
HIV-1 diversity can be attributed to:
- Error-prone reverse transcriptase enzyme
- Recombination of subtypes
- Cross-species transmission
Accurate quantitation is dependent upon a combination of:
- Primer Design
- Probe Design
- Cycling Conditions
| Primers and Probe are Targeted to the Integrase Region of the Polymerase Gene |
The integrase region of the polymerase gene is a conserved region of the HIV-1 genome.
| Partially Double Stranded Probe Design |
In the absence of target, the probe hybridizes to the quencher digoneucleotide, preventing fluorescent signal generation.
In the presence of target, the probe prefers to hybridize with the target sequence, disassociating from the quencher oligonucleotide and allowing fluorescent detection.
| Cycling Conditions: Low Temperature Read Cycles |
The cycling conditions for Abbott RealTime HIV-1 encompass a low temperature read cycle. This read cycle allows the probe to tolerate mismatches more effectively than a probe required to bind during the extension phase.
RealTime HIV-1 Qualitative
INTENDED USE
The Abbott RealTime HIV-1 assay is an in vitro reverse transcription-polymerase chain reaction (RT-PCR) assay for the quantitation of Human Immunodeficiency Virus type 1 (HIV-1) on the automated m2000 System in human plasma from HIV-1 infected individuals over the range of 40 to 10,000,000 copies/mL. The Abbott RealTime HIV-1 assay is intended for use in conjunction with clinical presentation and other laboratory markers for disease prognosis and for use as an aid in assessing viral response to antiretroviral treatment as measured by changes in plasma HIV-1 RNA levels. This assay is not intended to be used as a donor screening test for HIV-1 or as a diagnostic test to confirm the presence of HIV-1 infection.
Limitations
- Human plasma specimens collected in ACD-A or EDTA tubes may be used.
- A specimen with a result of "Target not detected" cannot be presumed to be negative for HIV-1 RNA.
- As with any diagnostic test, results from the Abbott RealTime HIV-1 assay should be interpreted in conjunction with other clinical and laboratory findings.
CAUTION: United States Federal law restricts this device to sale and distribution to or on the order of a physician or to a clinical laboratory; and use is restricted to, by, or on the order of a physician.
