This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by de Baar, M. P.
Right arrow Articles by de Ronde, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by de Baar, M. P.
Right arrow Articles by de Ronde, A.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, April 2001, p. 1378-1384, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1378-1384.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Single Rapid Real-Time Monitored Isothermal RNA Amplification Assay for Quantification of Human Immunodeficiency Virus Type 1 Isolates from Groups M, N, and O

Michel P. de Baar,1,* Maaike W. van Dooren,2 Esther de Rooij,1,2 Margreet Bakker,1,3 Bob van Gemen,2 Jaap Goudsmit,1,3 and Anthony de Ronde2

Department of Human Retrovirology, Academic Medical Center, University of Amsterdam,1 PrimaGen,2 and Amsterdam Institute of Viral Genomics,3 Amsterdam, The Netherlands

Received 7 September 2000/Returned for modification 6 November 2000/Accepted 26 January 2001

Because human immunodeficiency virus type 1 (HIV-1) subtypes and circulating recombinant forms (CRFs) are spreading rapidly worldwide and are becoming less confined to a geographical area, RNA assays that can detect and quantify all HIV-1 isolates reliably are in demand. We have developed a fast, real-time monitored RNA assay based on an isothermal nucleic acid sequence-based amplification technology that amplifies a part of the long terminal repeat region of the HIV-1 genome. Real-time detection was possible due to the addition of molecular beacons to the amplification reaction that was monitored in a fluorimeter with a thermostat. The lower level of detection of the assay was 10 HIV-1 RNA molecules per reaction, and the lower level of quantification was 100 copies of HIV-1 RNA with a dynamic range of linear quantification between 102 and 107 RNA molecules. All HIV-1 groups, subtypes, and CRFs could be detected and quantified with equal efficiency, including the group N isolate YBF30 and the group O isolate ANT70. To test the clinical utility of the assay, a series of 62 serum samples containing viruses that encompassed subtypes A through G and CRFs AE and AG of HIV-1 group M were analyzed, and these results were compared to the results of a commercially available assay. This comparison showed that the quantification results correlated highly (R2 = 0.735) for those subtypes that could be well quantified by both assays (subtypes B, C, D, and F), whereas improved quantification was obtained for subtypes A and G and CRFs AE and AG. A retrospective study with six individuals infected with either a subtype A, B, C, or D or an AG isolate of HIV-1 group M, who were treated with highly active antiretroviral therapy, revealed that the assay was well suited to the monitoring of therapy effects. In conclusion, the newly developed real-time monitored HIV-1 assay is a fast and sensitive assay with a large dynamic range of quantification and is suitable for quantification of most if not all subtypes and groups of HIV-1.

* Corresponding author. Mailing address: Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: 31-20-566 6780. Fax: 31-20-691 6531. E-mail: M.P.deBaar{at}amc.uva.nl.

Journal of Clinical Microbiology, April 2001, p. 1378-1384, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1378-1384.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Loens, K., Beck, T., Ursi, D., Overdijk, M., Sillekens, P., Goossens, H., Ieven, M. (2008). Development of Real-Time Multiplex Nucleic Acid Sequence-Based Amplification for Detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella spp. in Respiratory Specimens. J. Clin. Microbiol. 46: 185-191 [Abstract] [Full Text]  
  • Schutten, M., Peters, D., Back, N. K. T., Beld, M., Beuselinck, K., Foulongne, V., Geretti, A.-M., Pandiani, L., Tiemann, C., Niesters, H. G. M. (2007). Multicenter Evaluation of the New Abbott RealTime Assays for Quantitative Detection of Human Immunodeficiency Virus Type 1 and Hepatitis C Virus RNA. J. Clin. Microbiol. 45: 1712-1717 [Abstract] [Full Text]  
  • Ayele, W., Schuurman, R., Messele, T., Dorigo-Zetsma, W., Mengistu, Y., Goudsmit, J., Paxton, W. A., de Baar, M. P., Pollakis, G. (2007). Use of Dried Spots of Whole Blood, Plasma, and Mother's Milk Collected on Filter Paper for Measurement of Human Immunodeficiency Virus Type 1 Burden. J. Clin. Microbiol. 45: 891-896 [Abstract] [Full Text]  
  • Timmermans, E. C., Tebas, P., Ruiter, J. P.N., Wanders, R. J.A., de Ronde, A., de Baar, M. P. (2006). Real-Time Nucleic Acid Sequence-Based Amplification Assay to Quantify Changes in Mitochondrial DNA Concentrations in Cell Cultures and Blood Cells from HIV-Infected Patients Receiving Antiviral Therapy. Clin. Chem. 52: 979-987 [Abstract] [Full Text]  
  • Loens, K., Beck, T., Goossens, H., Ursi, D., Overdijk, M., Sillekens, P., Ieven, M. (2006). Development of Conventional and Real-Time Nucleic Acid Sequence-Based Amplification Assays for Detection of Chlamydophila pneumoniae in Respiratory Specimens. J. Clin. Microbiol. 44: 1241-1244 [Abstract] [Full Text]  
  • Muller, Z., Stelzl, E., Bozic, M., Haas, J., Marth, E., Kessler, H. H. (2004). Evaluation of Automated Sample Preparation and Quantitative PCR LCx Assay for Determination of Human Immunodeficiency Virus Type 1 RNA. J. Clin. Microbiol. 42: 1439-1443 [Abstract] [Full Text]  
  • Ayele, W., Pollakis, G., Abebe, A., Fisseha, B., Tegbaru, B., Tesfaye, G., Mengistu, Y., Wolday, D., van Gemen, B., Goudsmit, J., Dorigo-Zetsma, W., de Baar, M. P. (2004). Development of a Nucleic Acid Sequence-Based Amplification Assay That Uses gag-Based Molecular Beacons To Distinguish between Human Immunodeficiency Virus Type 1 Subtype C and C' Infections in Ethiopia. J. Clin. Microbiol. 42: 1534-1541 [Abstract] [Full Text]  
  • Prado, J. G., Shintani, A., Bofill, M., Clotet, B., Ruiz, L., Martinez-Picado, J. (2004). Lack of Longitudinal Intrapatient Correlation between p24 Antigenemia and Levels of Human Immunodeficiency Virus (HIV) Type 1 RNA in Patients with Chronic HIV Infection during Structured Treatment Interruptions. J. Clin. Microbiol. 42: 1620-1625 [Abstract] [Full Text]  
  • Antunes, R., Figueiredo, S., Bartolo, I., Pinheiro, M., Rosado, L., Soares, I., Lourenco, H., Taveira, N. (2003). Evaluation of the Clinical Sensitivities of Three Viral Load Assays with Plasma Samples from a Pediatric Population Predominantly Infected with Human Immunodeficiency Virus Type 1 Subtype G and BG Recombinant Forms. J. Clin. Microbiol. 41: 3361-3367 [Abstract] [Full Text]  
  • Marzio, G., Vink, M., Verhoef, K., de Ronde, A., Berkhout, B. (2002). Efficient Human Immunodeficiency Virus Replication Requires a Fine-Tuned Level of Transcription. J. Virol. 76: 3084-3088 [Abstract] [Full Text]  
  • de Baar, M. P., Timmermans, E. C., Bakker, M., de Rooij, E., van Gemen, B., Goudsmit, J. (2001). One-Tube Real-Time Isothermal Amplification Assay To Identify and Distinguish Human Immunodeficiency Virus Type 1 Subtypes A, B, and C and Circulating Recombinant Forms AE and AG. J. Clin. Microbiol. 39: 1895-1902 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»