Abbott RealTime HIV-1 m2000rt viral load testing: Manual extraction versus the automated m2000sp extraction

doi:10.1016/j.jviromet.2010.12.006

Journal of Virological Methods

Volume 172, Issues 1–2, March 2011, Pages 78-80

https://doi.org/10.1016/j.jviromet.2010.12.006 Get rights and content

Abstract

The Abbott RealTime HIV-1 assay is a real-time nucleic acid amplification assay available for HIV-1 viral load quantitation. The assay has a platform for automated extraction of viral RNA from plasma or dried blood spot samples, and an amplification platform with real time fluorescent detection. Overall, this study found no clinically relevant differences in viral load, if samples were extracted manually.

Section snippets

Acknowledgments

The authors wish to thank Lara Noble, Eveline Akkers and Kirthi Hira for their assistinace in sample and assay preparations conducted in Johannesburg, South Africa, the staff and patients of the Kilimanjaro Christian Medical Centre Infectious Diseases Clinic, and the staff and clients of Kikundi cha Wanawake Kilimanjaro Kupambana na UKIMWI (KIWAKKUKI; Women Against AIDS in Kilimanjaro) for their participation. The authors thank Ekyafyose E. Kimaro, Julitha Kimbi, Devotha Lyimo, and Editha Mushi

References (7)

J.M. Bland et al.
Comparing methods of measurement: why plotting difference against standard method is misleading
Lancet
(1995)
J.A. Crump et al.
Evaluation of the Abbott m2000rt RealTime HIV-1 assay with manual sample preparation compared with the ROCHE COBAS AmpliPrep/AMPLICOR HIV-1 MONITOR v1.5 using specimens from East Africa
J. Virol. Methods
(2009)
G. Yilmaz
Diagnosis of HIV infection and laboratory monitoring of its therapy
J. Clin. Virol.
(2001)

There are more references available in the full text version of this article.

Cited by (21)

Cost-effective HIV-1 virological monitoring in resource-limited settings using a modified commercially available qPCR RNA assay
2017, Journal of Virological Methods
Citation Excerpt :
To minimize expenses, we carried out the modified assay manually, i.e. without using the Abbott’s m2000sp automated system for RNA extraction and qPCR plate setup. Previous studies found good agreement between results from manual versus automated Abbott assay (Scott et al., 2011), as well as between the manual Abbott assay and qPCR-based assays from different suppliers (Crump et al., 2009). An additional option for cost reduction is to run screening assays on pooled samples, rather than individual samples, a strategy that has been successfully adapted to virological monitoring for identification of ART failure, and shown to effectively reduce monitoring costs (Kim et al., 2014; May et al., 2010).
Virological monitoring through plasma viral load (PVL) quantification is essential for clinical management of HIV patients undergoing antiretroviral treatment (ART), and for detecting treatment failure. Quantitative PCR (qPCR)-based tests are the gold standard for measuring PVL. Largely because of their high cost, however, implementation of these tests in low- and middle-income countries fails to cover the testing demand. In this study, we aimed at reducing the running cost of the commercially available Abbott RealTime™ HIV-1 assay by minimizing the reagent consumption. To this end, a modified version of the assay was obtained by reducing the assay’s reagents volume to about a half, and validated using a panel of 104 plasma samples. Compared to the standard version, the modified Abbott assay allowed for a 50% reduction in running costs. At the same time, it showed a 100% concordance in identifying samples with detectable viral load, strong correlation (Pearson’s r = 0.983, P < 0.0001), and a high agreement between PVL values (mean percent difference between PVL values ± standard deviation = 0.76 ± 3.18%). In detecting viral failure (PVL > 1000 copies mL⁻¹), the modified assay showed a sensitivity of 94.6%, a specificity of 93.8%, and a negative and positive predictive values of 93.8% and 94.6%, respectively. The modified assay therefore reliably quantifies PVL, predicts viral failure, and allows for a ca. 50% reduction in the assay’s running costs. It may thus be implemented as an ART monitoring tool in resource-limited settings and for research purposes.
Utility of rapid antibody tests to exclude HIV-1 infection among infants and children aged <18 months in a low-resource setting
2012, Journal of Clinical Virology
Citation Excerpt :
Medical history, physical examination, and a blood culture were also obtained.8 Children under the age of 18 months with two positive RT had CD4+ T-lymphocyte count and percentage along with serum or plasma HIV-1 RNA measured by NAAT (Abbott Real-Time m2000 System, Abbott Molecular, Illinois, USA).10,11 Children with discordant RT had HIV-1 enzyme linked immunosorbent assay (ELISA) (Vironistika Uniform II Plus-O Test, bio-Mérieux, NC, USA), but no further testing if negative on ELISA.
Excluding HIV infection among infants and young children in resource-poor settings where nucleic acid amplification tests (NAAT) are not routinely available remains a considerable challenge.
To assess the performance of two rapid HIV antibody tests (RT) used alone and in parallel for excluding HIV infection among acutely ill infants and children <18 months in comparison to NAAT in a region where maternal HIV prevalence was approximately 7%.
Infants and children ≥2 < 18 months admitted to hospital with an acute febrile illness had two rapid antibody tests in parallel, with single and parallel results subsequently compared against NAAT.
HIV prevalence among 1602 enrolled infants was 3.4%. All 1526 infants with 2 negative RT were HIV negative by NAAT. All 46 infants with 2 positive RT were HIV positive by NAAT. The overall specificity of two rapid tests for excluding HIV infection was 99.5%. Sensitivity and specificity were ≥99% and >98%, respectively, across all age brackets ≥2 < 18 months. Overall sensitivity and specificity for a single RT was 98.2% and 99%, respectively, for Determine, and 85.5% and 99.6%, respectively, for Capillus.
In a setting with a maternal HIV prevalence rate of <10%, a single negative RT had excellent specificity and two negative RT performed in parallel had a perfect negative predictive value for HIV infection among acutely ill patients <18 months of age. In this and similar settings, RT could assist with excluding HIV infection with much lower complexity and cost than NAAT.
A new highly automated extraction system for quantitative real-time PCRs from whole blood samples: Routine monitoring of opportunistic infections in immunosuppressed patients
2012, Journal of Clinical Virology
Citation Excerpt :
But this, in turn, has dramatically increased the number of routine tests performed by virology departments signalling the need for fully automated extraction instruments. Currently plasma samples are used routinely for quantifying HIV,3,4 HCV,5 and HBV virus nucleic acids with high throughput automated platforms. The samples used for molecular assays of other viruses such as cytomegalovirus (CMV), Epstein–Barr virus (EBV), and BK virus may vary from serum and plasma to whole blood, depending on local preference.
Rapid, high throughput extraction systems are needed to monitor viral infections in immunosuppressed patients.
Evaluate the performance of the MagNA Pure 96™ extraction system, and compare it to the COBAS Ampliprep™ for quantitative real-time PCR from whole blood samples.
Compare the MagNA Pure LC™, COBAS Ampliprep™ and MagNA Pure 96™ using ten-fold dilutions of blood samples containing cytomegalovirus. Evaluate analytical performances of the MagNA Pure 96™ from test samples containing cytomegalovirus. Evaluate clinical performances from 209 blood samples collected prospectively, extracted with the COBAS Ampliprep™ and the MagNA Pure 96™ systems and tested for cytomegalovirus, Epstein–Barr, BK and JC viruses.
All three extraction systems gave similar results with dilutions of a cytomegalovirus-positive sample. Analytical tests showed that the limit of detection was 500 copies/ml, specificity was 100%, with no cross-contamination. Quantification was linear from 3.0 to 6.0 log₁₀ copies/ml. Intra-assay variation was 8.3–0.9% and inter-assay variation 8.8–5.2%. Clinical specimens extracted with the MagNA Pure 96™ and COBAS Ampliprep™ instruments agreed well for cytomegalovirus (r = 0.54; p = 0.07), Epstein–Barr virus (0.69; p = 0.0005) and BK virus (0.85; p = 0.01). All 55 samples were negative for JC virus. Mean loads were similar for cytomegalovirus (0.17 log₁₀ copies/ml) and BK virus (−0.24 log₁₀ copies/ml) while that of Epstein–Barr virus was slightly lower (1.02 log₁₀ copies/ml).
The MagNA Pure 96™ instrument is an easy-to-use, reliable high throughput platform for extracting nucleic acid from clinical whole blood specimens.
Suitability of an open automated nucleic acid extractor for high-throughput plasma HIV-1 RNA quantitation in Gabon (Central Africa)
2012, Journal of Virological Methods
Nucleic acid extraction using the open automated EZ1 (Qiagen) instrument, in combination with the Generic HIV Viral Load assay, gave highly concordant HIV-1 RNA viral load results among 181 Gabonese subjects infected with HIV-1, compared to those obtained when performing a manual extraction. Since people living with HIV-1 are being treated with antiretrovirals in Gabon, this automated extraction technique represents an excellent technical method for high-throughput monitoring of HIV-1 RNA viral load.
Quantification of serum HDV RNA by Robogene 2.0 in HDV patients is significantly influenced by the extraction methods
2024, Liver International
Changes in Serum Levels of IL-6, TNF-α and CRP in People Living with HIV Following Initiation of Antiretroviral Therapy — A Longitudinal Cohort Study from a Tertiary Hospital In Eastern India
2023, Journal of the Indian Medical Association

View all citing articles on Scopus

View full text

Short communicationAbbott RealTime HIV-1 m2000rt viral load testing: Manual extraction versus the automated m2000sp extraction

Abstract

Section snippets

Acknowledgments

Lancet

J. Virol. Methods

J. Clin. Virol.

Short communication
Abbott RealTime HIV-1 m2000rt viral load testing: Manual extraction versus the automated m2000sp extraction