One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil
ABSTRACT The Brazilian Public Health Service provides freely αPEG-IFN to treat patients infected with HCV. The primary goal of HCV therapy is the long-term elimination of HCV from the blood to reduce the risk of HCV associated complications and death. Patient viremia affects the treatment duration and response, thus influencing clinical decisions. We developed a high-throughput method to perform the quantification of RNA hepatitis C virus (HCV) virus load in plasma samples to monitor patients under treatment. The method is based on a duplex detection, in a one-step real-time RT-PCR assay and it has been validated according to the rules established by the official Brazilian regulatory agency (ANVISA). This new method was compared to a commercial kit (Cobas/Taqman HCV Test v2.0 - Roche), showing virus load results with significant correlation between them (p= 0,012) using commercial and clinical panels. In addition, 611 samples from patients treated with peguilated alfa-interferon (αPEG-IFN) from different regions of Brazil were analyzed. Our one-step real-time RT-PCR assay demonstrated good performance in viral load measurement and in treatment course monitoring, with acceptable sensitivity and specificity values.
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| Language: | English |
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Brazilian Society of Infectious Diseases
2018
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| Online Access: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702018000500418 |
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oai:scielo:S1413-867020180005004182018-12-14One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in BrazilAndrade,ElisabeteRocha,DanieleFontana-Maurell,MarcelaCosta,ElaineRibeiro,Marisade Godoy,Daniela TupyFerreira,Antonio G.P.Tanuri,AmilcarAlvarez,PatríciaBrindeiro,Rodrigo HCV viral load Real Time PCR monitoring patients ABSTRACT The Brazilian Public Health Service provides freely αPEG-IFN to treat patients infected with HCV. The primary goal of HCV therapy is the long-term elimination of HCV from the blood to reduce the risk of HCV associated complications and death. Patient viremia affects the treatment duration and response, thus influencing clinical decisions. We developed a high-throughput method to perform the quantification of RNA hepatitis C virus (HCV) virus load in plasma samples to monitor patients under treatment. The method is based on a duplex detection, in a one-step real-time RT-PCR assay and it has been validated according to the rules established by the official Brazilian regulatory agency (ANVISA). This new method was compared to a commercial kit (Cobas/Taqman HCV Test v2.0 - Roche), showing virus load results with significant correlation between them (p= 0,012) using commercial and clinical panels. In addition, 611 samples from patients treated with peguilated alfa-interferon (αPEG-IFN) from different regions of Brazil were analyzed. Our one-step real-time RT-PCR assay demonstrated good performance in viral load measurement and in treatment course monitoring, with acceptable sensitivity and specificity values.info:eu-repo/semantics/openAccessBrazilian Society of Infectious DiseasesBrazilian Journal of Infectious Diseases v.22 n.5 20182018-10-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702018000500418en10.1016/j.bjid.2018.08.003 |
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| author |
Andrade,Elisabete Rocha,Daniele Fontana-Maurell,Marcela Costa,Elaine Ribeiro,Marisa de Godoy,Daniela Tupy Ferreira,Antonio G.P. Tanuri,Amilcar Alvarez,Patrícia Brindeiro,Rodrigo |
| spellingShingle |
Andrade,Elisabete Rocha,Daniele Fontana-Maurell,Marcela Costa,Elaine Ribeiro,Marisa de Godoy,Daniela Tupy Ferreira,Antonio G.P. Tanuri,Amilcar Alvarez,Patrícia Brindeiro,Rodrigo One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil |
| author_facet |
Andrade,Elisabete Rocha,Daniele Fontana-Maurell,Marcela Costa,Elaine Ribeiro,Marisa de Godoy,Daniela Tupy Ferreira,Antonio G.P. Tanuri,Amilcar Alvarez,Patrícia Brindeiro,Rodrigo |
| author_sort |
Andrade,Elisabete |
| title |
One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil |
| title_short |
One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil |
| title_full |
One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil |
| title_fullStr |
One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil |
| title_full_unstemmed |
One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil |
| title_sort |
one-step real-time pcr assay for detection and quantification of rna hcv to monitor patients under treatment in brazil |
| description |
ABSTRACT The Brazilian Public Health Service provides freely αPEG-IFN to treat patients infected with HCV. The primary goal of HCV therapy is the long-term elimination of HCV from the blood to reduce the risk of HCV associated complications and death. Patient viremia affects the treatment duration and response, thus influencing clinical decisions. We developed a high-throughput method to perform the quantification of RNA hepatitis C virus (HCV) virus load in plasma samples to monitor patients under treatment. The method is based on a duplex detection, in a one-step real-time RT-PCR assay and it has been validated according to the rules established by the official Brazilian regulatory agency (ANVISA). This new method was compared to a commercial kit (Cobas/Taqman HCV Test v2.0 - Roche), showing virus load results with significant correlation between them (p= 0,012) using commercial and clinical panels. In addition, 611 samples from patients treated with peguilated alfa-interferon (αPEG-IFN) from different regions of Brazil were analyzed. Our one-step real-time RT-PCR assay demonstrated good performance in viral load measurement and in treatment course monitoring, with acceptable sensitivity and specificity values. |
| publisher |
Brazilian Society of Infectious Diseases |
| publishDate |
2018 |
| url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702018000500418 |
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