Multiplex real-time quantitative RT-PCR assay for hepatitis B virus, hepatitis C virus, and human immunodeficiency virus type 1.
Identifieur interne : 000F22 ( Main/Curation ); précédent : 000F21; suivant : 000F23Multiplex real-time quantitative RT-PCR assay for hepatitis B virus, hepatitis C virus, and human immunodeficiency virus type 1.
Auteurs : Daniel Candotti [Royaume-Uni] ; Jillian Temple ; Shirley Owusu-Ofori ; Jean-Pierre AllainSource :
- Journal of virological methods [ 0166-0934 ] ; 2004.
Descripteurs français
- KwdFr :
- ADN viral (génétique), ADN viral (sang), ARN viral (génétique), ARN viral (sang), Donneurs de sang, Ghana, Génotype, Hepacivirus (génétique), Hepacivirus (isolement et purification), Humains, RT-PCR (), Séquence nucléotidique, VIH-1 (Virus de l'Immunodéficience Humaine de type 1) (génétique), VIH-1 (Virus de l'Immunodéficience Humaine de type 1) (isolement et purification), Virologie (), Virus de l'hépatite B (génétique), Virus de l'hépatite B (isolement et purification).
- MESH :
- génétique : ADN viral, ARN viral, Hepacivirus, VIH-1 (Virus de l'Immunodéficience Humaine de type 1), Virus de l'hépatite B.
- isolement et purification : Hepacivirus, VIH-1 (Virus de l'Immunodéficience Humaine de type 1), Virus de l'hépatite B.
- sang : ADN viral, ARN viral.
- Donneurs de sang, Ghana, Génotype, Humains, RT-PCR, Séquence nucléotidique, Virologie.
- Wicri :
- geographic : Ghana.
English descriptors
- KwdEn :
- Base Sequence, Blood Donors, DNA, Viral (blood), DNA, Viral (genetics), Genotype, Ghana, HIV-1 (genetics), HIV-1 (isolation & purification), Hepacivirus (genetics), Hepacivirus (isolation & purification), Hepatitis B virus (genetics), Hepatitis B virus (isolation & purification), Humans, RNA, Viral (blood), RNA, Viral (genetics), Reverse Transcriptase Polymerase Chain Reaction (methods), Virology (methods).
- MESH :
- chemical , blood : DNA, Viral, RNA, Viral.
- chemical , genetics : DNA, Viral, RNA, Viral.
- geographic : Ghana.
- genetics : HIV-1, Hepacivirus, Hepatitis B virus.
- isolation & purification : HIV-1, Hepacivirus, Hepatitis B virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction, Virology.
- Base Sequence, Blood Donors, Genotype, Humans.
Abstract
A multiplex real-time quantitative reverse transcription (RT)-PCR assay was developed for simultaneous detection, identification and quantification of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) in plasma or serum samples. Genomic amplification of one virus was unaffected by the simultaneous amplification of the other two. Competition between HCV and HIV-1 amplifications slightly affected the yield of HIV-1 amplification. However, quantitation was possible when a single virus was present. The 95% detection limits were 30, 167 and 680IU/ml for HBV DNA, HCV RNA and HIV-1 RNA, respectively. The multiplex assay detected with similar efficiency strains of HBV genotypes A-F, HCV genotypes 1-6, and HIV-1 subtypes A-G. Applied to 267 pools of 10 plasmas from blood donors, multiplex screening indicated that the assay was reproducible, sensitive, and specific. This assay has the potential to be used for large-scale nucleic acid testing (NAT) of blood donations.
DOI: 10.1016/j.jviromet.2004.01.017
PubMed: 15158067
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pubmed:15158067Le document en format XML
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<author><name sortKey="Owusu Ofori, Shirley" sort="Owusu Ofori, Shirley" uniqKey="Owusu Ofori S" first="Shirley" last="Owusu-Ofori">Shirley Owusu-Ofori</name>
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<series><title level="j">Journal of virological methods</title>
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<term>Blood Donors</term>
<term>DNA, Viral (blood)</term>
<term>DNA, Viral (genetics)</term>
<term>Genotype</term>
<term>Ghana</term>
<term>HIV-1 (genetics)</term>
<term>HIV-1 (isolation & purification)</term>
<term>Hepacivirus (genetics)</term>
<term>Hepacivirus (isolation & purification)</term>
<term>Hepatitis B virus (genetics)</term>
<term>Hepatitis B virus (isolation & purification)</term>
<term>Humans</term>
<term>RNA, Viral (blood)</term>
<term>RNA, Viral (genetics)</term>
<term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term>
<term>Virology (methods)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>ADN viral (génétique)</term>
<term>ADN viral (sang)</term>
<term>ARN viral (génétique)</term>
<term>ARN viral (sang)</term>
<term>Donneurs de sang</term>
<term>Ghana</term>
<term>Génotype</term>
<term>Hepacivirus (génétique)</term>
<term>Hepacivirus (isolement et purification)</term>
<term>Humains</term>
<term>RT-PCR ()</term>
<term>Séquence nucléotidique</term>
<term>VIH-1 (Virus de l'Immunodéficience Humaine de type 1) (génétique)</term>
<term>VIH-1 (Virus de l'Immunodéficience Humaine de type 1) (isolement et purification)</term>
<term>Virologie ()</term>
<term>Virus de l'hépatite B (génétique)</term>
<term>Virus de l'hépatite B (isolement et purification)</term>
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<term>RNA, Viral</term>
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<term>RNA, Viral</term>
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<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>HIV-1</term>
<term>Hepacivirus</term>
<term>Hepatitis B virus</term>
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<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN viral</term>
<term>ARN viral</term>
<term>Hepacivirus</term>
<term>VIH-1 (Virus de l'Immunodéficience Humaine de type 1)</term>
<term>Virus de l'hépatite B</term>
</keywords>
<keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>HIV-1</term>
<term>Hepacivirus</term>
<term>Hepatitis B virus</term>
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<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Hepacivirus</term>
<term>VIH-1 (Virus de l'Immunodéficience Humaine de type 1)</term>
<term>Virus de l'hépatite B</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>Virology</term>
</keywords>
<keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>ADN viral</term>
<term>ARN viral</term>
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<term>Genotype</term>
<term>Humans</term>
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<term>Ghana</term>
<term>Génotype</term>
<term>Humains</term>
<term>RT-PCR</term>
<term>Séquence nucléotidique</term>
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<front><div type="abstract" xml:lang="en">A multiplex real-time quantitative reverse transcription (RT)-PCR assay was developed for simultaneous detection, identification and quantification of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) in plasma or serum samples. Genomic amplification of one virus was unaffected by the simultaneous amplification of the other two. Competition between HCV and HIV-1 amplifications slightly affected the yield of HIV-1 amplification. However, quantitation was possible when a single virus was present. The 95% detection limits were 30, 167 and 680IU/ml for HBV DNA, HCV RNA and HIV-1 RNA, respectively. The multiplex assay detected with similar efficiency strains of HBV genotypes A-F, HCV genotypes 1-6, and HIV-1 subtypes A-G. Applied to 267 pools of 10 plasmas from blood donors, multiplex screening indicated that the assay was reproducible, sensitive, and specific. This assay has the potential to be used for large-scale nucleic acid testing (NAT) of blood donations.</div>
</front>
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