Detection of HIV-1 RNA in plasma and serum samples using the NASBA amplification system compared to RNA-PCR
Identifieur interne : 001352 ( Main/Exploration ); précédent : 001351; suivant : 001353Detection of HIV-1 RNA in plasma and serum samples using the NASBA amplification system compared to RNA-PCR
Auteurs : Anne-Mieke Vandamme [Belgique] ; Sonia Van Dooren [Belgique] ; Wessel Kok [Pays-Bas] ; Patrick Goubau [Belgique] ; Katrien Fransen [Belgique] ; Tim Kievits [Pays-Bas] ; Jean-Claude Schmit [Belgique] ; Erik De Clercq [Belgique] ; Jan Desmyter [Belgique]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 1995.
English descriptors
- KwdEn :
- Amplification, Amplification products, Amplification system, Antisense, Assay, Belgian aids reference laboratories, Ccid, Cells lysed, Control plasmid, Detection limit, Dextran sulfate, Dilution series, Disease stage, Elution buffer, European community, European seropositive, Extraction method, Gemen, Healthy seropositives, Horse radish peroxidase, Human immunodeficiency virus type, Hxb2, Internal control, Kenyan seronegative, Kievits, Latent stage, Monitoring changes, Nasba, Nasba amplification system, Nasba product, Nasba system, Nasba test, Negative controls, Nucleic acids, Nucleotide sequence, Organon teknika, Other results, Patient samples, Plasma, Plasma samples, Plasmid, Plasmid pgem3, Polymerase, Polymerase chain reaction, Positive result, Primary infection, Primer, Primer sets, Promoter sequence, Sensitive techniques, Seronegative plasma, Seropositives, Serum samples, Silica particles, Simple method, Spiked plasma, Standard curve, System control, Vandamme, Viral, Viral load, Virological, Virological methods, Virus stock.
- Teeft :
- Amplification, Amplification products, Amplification system, Antisense, Assay, Belgian aids reference laboratories, Ccid, Cells lysed, Control plasmid, Detection limit, Dextran sulfate, Dilution series, Disease stage, Elution buffer, European community, European seropositive, Extraction method, Gemen, Healthy seropositives, Horse radish peroxidase, Human immunodeficiency virus type, Hxb2, Internal control, Kenyan seronegative, Kievits, Latent stage, Monitoring changes, Nasba, Nasba amplification system, Nasba product, Nasba system, Nasba test, Negative controls, Nucleic acids, Nucleotide sequence, Organon teknika, Other results, Patient samples, Plasma, Plasma samples, Plasmid, Plasmid pgem3, Polymerase, Polymerase chain reaction, Positive result, Primary infection, Primer, Primer sets, Promoter sequence, Sensitive techniques, Seronegative plasma, Seropositives, Serum samples, Silica particles, Simple method, Spiked plasma, Standard curve, System control, Vandamme, Viral, Viral load, Virological, Virological methods, Virus stock.
Abstract
The presence of HIV-1 RNA in the plasma and serum of European and African patients was monitored using RNA-polymerase chain reaction (RNA-PCR) and the new isothermal NASBA nucleic acid amplification system encompassing a gel-based detection assay (ELGA). Identical RNA extraction procedures, provided by the NASBA amplification system, were used for both methods. The detection limit for HIV-1 RNA, measured on a 10-fold dilution series of spiked HIVIIIB in negative plasma, was about 0.05 CCID50 per test for both methods. Both NASBA and RNA-PCR were more sensitive than a p24 assay for the detection of circulating HIV-1 virus in blood: 17 of the 34 (50%) p24 antigen-tested seropositives were p24-positive while 32 (94%) were positive by NASBA and 30 (88%) by RNA-PCR. Among the 45 seropositives, 34 of which were tested for p24 antigen, 43 (96%) were positive by NASBA and 41 (91%) by RNA-PCR. Almost all seropositives had a detectable viral load in 100 μl plasma. Lower viral loads were only encountered in some healthy seropositives with a higher CD4 count. There was no cross-reactivity with HIV-2 or HTLV-I with both the RNA-PCR and NASBA. The extraction method used permitted the detection of HIV-1 RNA equally well in serum and in plasma with heparin or EDTA.
Url:
DOI: 10.1016/0166-0934(94)00151-6
Affiliations:
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xml:lang="fr">Belgique</country><wicri:regionArea>Rega Institute for Medical Research and University Hospitals, Katholieke Universiteit Leuven, B-3000 Leuven</wicri:regionArea><wicri:noRegion>B-3000 Leuven</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="ISSN">0166-0934</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1995">1995</date><biblScope unit="volume">52</biblScope><biblScope unit="issue">1–2</biblScope><biblScope unit="page" from="121">121</biblScope><biblScope unit="page" to="132">132</biblScope></imprint><idno type="ISSN">0166-0934</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amplification</term><term>Amplification products</term><term>Amplification system</term><term>Antisense</term><term>Assay</term><term>Belgian aids reference laboratories</term><term>Ccid</term><term>Cells lysed</term><term>Control plasmid</term><term>Detection limit</term><term>Dextran sulfate</term><term>Dilution series</term><term>Disease stage</term><term>Elution buffer</term><term>European community</term><term>European seropositive</term><term>Extraction method</term><term>Gemen</term><term>Healthy seropositives</term><term>Horse radish peroxidase</term><term>Human immunodeficiency virus type</term><term>Hxb2</term><term>Internal control</term><term>Kenyan seronegative</term><term>Kievits</term><term>Latent stage</term><term>Monitoring changes</term><term>Nasba</term><term>Nasba amplification system</term><term>Nasba product</term><term>Nasba system</term><term>Nasba test</term><term>Negative controls</term><term>Nucleic acids</term><term>Nucleotide sequence</term><term>Organon teknika</term><term>Other results</term><term>Patient samples</term><term>Plasma</term><term>Plasma samples</term><term>Plasmid</term><term>Plasmid pgem3</term><term>Polymerase</term><term>Polymerase chain reaction</term><term>Positive result</term><term>Primary infection</term><term>Primer</term><term>Primer sets</term><term>Promoter sequence</term><term>Sensitive techniques</term><term>Seronegative plasma</term><term>Seropositives</term><term>Serum samples</term><term>Silica particles</term><term>Simple method</term><term>Spiked plasma</term><term>Standard curve</term><term>System control</term><term>Vandamme</term><term>Viral</term><term>Viral load</term><term>Virological</term><term>Virological methods</term><term>Virus stock</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Amplification</term><term>Amplification products</term><term>Amplification system</term><term>Antisense</term><term>Assay</term><term>Belgian aids reference laboratories</term><term>Ccid</term><term>Cells lysed</term><term>Control plasmid</term><term>Detection limit</term><term>Dextran sulfate</term><term>Dilution series</term><term>Disease stage</term><term>Elution buffer</term><term>European community</term><term>European seropositive</term><term>Extraction method</term><term>Gemen</term><term>Healthy seropositives</term><term>Horse radish peroxidase</term><term>Human immunodeficiency virus type</term><term>Hxb2</term><term>Internal control</term><term>Kenyan seronegative</term><term>Kievits</term><term>Latent stage</term><term>Monitoring changes</term><term>Nasba</term><term>Nasba amplification system</term><term>Nasba product</term><term>Nasba system</term><term>Nasba test</term><term>Negative controls</term><term>Nucleic acids</term><term>Nucleotide sequence</term><term>Organon teknika</term><term>Other results</term><term>Patient samples</term><term>Plasma</term><term>Plasma samples</term><term>Plasmid</term><term>Plasmid pgem3</term><term>Polymerase</term><term>Polymerase chain reaction</term><term>Positive result</term><term>Primary infection</term><term>Primer</term><term>Primer sets</term><term>Promoter sequence</term><term>Sensitive techniques</term><term>Seronegative plasma</term><term>Seropositives</term><term>Serum samples</term><term>Silica particles</term><term>Simple method</term><term>Spiked plasma</term><term>Standard curve</term><term>System control</term><term>Vandamme</term><term>Viral</term><term>Viral load</term><term>Virological</term><term>Virological methods</term><term>Virus stock</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The presence of HIV-1 RNA in the plasma and serum of European and African patients was monitored using RNA-polymerase chain reaction (RNA-PCR) and the new isothermal NASBA nucleic acid amplification system encompassing a gel-based detection assay (ELGA). Identical RNA extraction procedures, provided by the NASBA amplification system, were used for both methods. The detection limit for HIV-1 RNA, measured on a 10-fold dilution series of spiked HIVIIIB in negative plasma, was about 0.05 CCID50 per test for both methods. Both NASBA and RNA-PCR were more sensitive than a p24 assay for the detection of circulating HIV-1 virus in blood: 17 of the 34 (50%) p24 antigen-tested seropositives were p24-positive while 32 (94%) were positive by NASBA and 30 (88%) by RNA-PCR. Among the 45 seropositives, 34 of which were tested for p24 antigen, 43 (96%) were positive by NASBA and 41 (91%) by RNA-PCR. Almost all seropositives had a detectable viral load in 100 μl plasma. Lower viral loads were only encountered in some healthy seropositives with a higher CD4 count. There was no cross-reactivity with HIV-2 or HTLV-I with both the RNA-PCR and NASBA. The extraction method used permitted the detection of HIV-1 RNA equally well in serum and in plasma with heparin or EDTA.</div></front></TEI><affiliations><list><country><li>Belgique</li><li>Pays-Bas</li></country></list><tree><country name="Belgique"><noRegion><name sortKey="Vandamme, Anne Mieke" sort="Vandamme, Anne Mieke" uniqKey="Vandamme A" first="Anne-Mieke" last="Vandamme">Anne-Mieke Vandamme</name></noRegion><name sortKey="De Clercq, Erik" sort="De Clercq, Erik" uniqKey="De Clercq E" first="Erik" last="De Clercq">Erik De Clercq</name><name sortKey="Desmyter, Jan" sort="Desmyter, Jan" uniqKey="Desmyter J" first="Jan" last="Desmyter">Jan Desmyter</name><name sortKey="Fransen, Katrien" sort="Fransen, Katrien" uniqKey="Fransen K" first="Katrien" last="Fransen">Katrien Fransen</name><name sortKey="Goubau, Patrick" sort="Goubau, Patrick" uniqKey="Goubau P" first="Patrick" last="Goubau">Patrick Goubau</name><name sortKey="Schmit, Jean Claude" sort="Schmit, Jean Claude" uniqKey="Schmit J" first="Jean-Claude" last="Schmit">Jean-Claude Schmit</name><name sortKey="Van Dooren, Sonia" sort="Van Dooren, Sonia" uniqKey="Van Dooren S" first="Sonia" last="Van Dooren">Sonia Van Dooren</name></country><country name="Pays-Bas"><noRegion><name sortKey="Kok, Wessel" sort="Kok, Wessel" uniqKey="Kok W" first="Wessel" last="Kok">Wessel Kok</name></noRegion><name sortKey="Kievits, Tim" sort="Kievits, Tim" uniqKey="Kievits T" first="Tim" last="Kievits">Tim Kievits</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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