Highly sensitive plasma RNA quantification by real-time PCR in HIV-2 group A and group B infection
Copyright © 2013 Elsevier B.V. All rights reserved..
UNLABELLED: Plasma HIV-2 viral load has been reported as predictive of AIDS in HIV-2 infected patient but the lack of sensitivity of the current HIV2 viral load assay is a limitation for the monitoring of the HIV-2-infected patients.
OBJECTIVE: To validate a new quantification assay based on a synthetic HIV-2 RNA transcript and real-time PCR with primers and probes selected in the LTR region, together with high-performance reagents and a protective RNA carrier.
STUDY DESIGN: We quantified 23 HIV-2 group A and B supernatants and 58 plasma samples with our TAQMAN-PCR assay and compared the results to those of our previously published in a real time reference PCR performed onto Light Cycler technology, the LC-PCR with a detection of 2.0 log10 copies/ml.
RESULTS: The performance of TAQMAN-PCR was significantly improved, yielding a detection limit of 17 RNA copies/ml. There was a major difference (1-5 log10 copies/ml) between LC-PCR and TAQMAN-PCR values for HIV-2 group B supernatants. Twenty-six of 27 plasma samples with levels higher than 2.0 log10 copies/ml in LC-PCR were positive by TAQMAN-PCR. Ten of the 31 plasma samples below the LC-PCR detection limit were quantifiable with the TAQMAN-PCR.
CONCLUSIONS: The new primers and probe in the LTR region can circumvent HIV-2 diversity, making our method suitable for use in HIV-2 group B-infected patients. Use of a high-performance RT enzyme and RNA carrier protection contributed to improving the detection limit. This study confirms that plasma viral load is lower than 17 copies/ml in a large number of HIV-2-infected patients.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2013 |
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| Erschienen: |
2013 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:58 |
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| Enthalten in: |
Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology - 58(2013), 2 vom: 10. Okt., Seite 461-7 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Delarue, Séverine [VerfasserIn] |
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| Links: |
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| Themen: |
DNA Primers |
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| Anmerkungen: |
Date Completed 18.04.2014 Date Revised 10.12.2019 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jcv.2013.08.003 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM23061468X |
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| 100 | 1 | |a Delarue, Séverine |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Highly sensitive plasma RNA quantification by real-time PCR in HIV-2 group A and group B infection |
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| 500 | |a Date Completed 18.04.2014 | ||
| 500 | |a Date Revised 10.12.2019 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2013 Elsevier B.V. All rights reserved. | ||
| 520 | |a UNLABELLED: Plasma HIV-2 viral load has been reported as predictive of AIDS in HIV-2 infected patient but the lack of sensitivity of the current HIV2 viral load assay is a limitation for the monitoring of the HIV-2-infected patients | ||
| 520 | |a OBJECTIVE: To validate a new quantification assay based on a synthetic HIV-2 RNA transcript and real-time PCR with primers and probes selected in the LTR region, together with high-performance reagents and a protective RNA carrier | ||
| 520 | |a STUDY DESIGN: We quantified 23 HIV-2 group A and B supernatants and 58 plasma samples with our TAQMAN-PCR assay and compared the results to those of our previously published in a real time reference PCR performed onto Light Cycler technology, the LC-PCR with a detection of 2.0 log10 copies/ml | ||
| 520 | |a RESULTS: The performance of TAQMAN-PCR was significantly improved, yielding a detection limit of 17 RNA copies/ml. There was a major difference (1-5 log10 copies/ml) between LC-PCR and TAQMAN-PCR values for HIV-2 group B supernatants. Twenty-six of 27 plasma samples with levels higher than 2.0 log10 copies/ml in LC-PCR were positive by TAQMAN-PCR. Ten of the 31 plasma samples below the LC-PCR detection limit were quantifiable with the TAQMAN-PCR | ||
| 520 | |a CONCLUSIONS: The new primers and probe in the LTR region can circumvent HIV-2 diversity, making our method suitable for use in HIV-2 group B-infected patients. Use of a high-performance RT enzyme and RNA carrier protection contributed to improving the detection limit. This study confirms that plasma viral load is lower than 17 copies/ml in a large number of HIV-2-infected patients | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Validation Study | |
| 650 | 4 | |a HIV-2 | |
| 650 | 4 | |a Natural history | |
| 650 | 4 | |a Viral load | |
| 650 | 7 | |a DNA Primers |2 NLM | |
| 650 | 7 | |a Oligonucleotide Probes |2 NLM | |
| 650 | 7 | |a RNA, Viral |2 NLM | |
| 700 | 1 | |a Didier, Emmanuelle |e verfasserin |4 aut | |
| 700 | 1 | |a Damond, Florence |e verfasserin |4 aut | |
| 700 | 1 | |a Ponscarme, Diane |e verfasserin |4 aut | |
| 700 | 1 | |a Brengle-Pesce, Karen |e verfasserin |4 aut | |
| 700 | 1 | |a Resche-Rigon, Matthieu |e verfasserin |4 aut | |
| 700 | 1 | |a Vray, Muriel |e verfasserin |4 aut | |
| 700 | 1 | |a Gueudin, Marie |e verfasserin |4 aut | |
| 700 | 1 | |a Simon, François |e verfasserin |4 aut | |
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