In-house quantitative real-time PCR for the diagnosis of hepatitis B virus and hepatitis C virus infections

Zauli, Danielle Alves Gomes; Menezes, Carla Lisandre Paula de; Oliveira, Cristiane Lommez de; Mateo, Elvis Cristian Cueva; Ferreira, Alessandro Clayton de Souza

doi:10.1016/j.bjm.2016.07.008

Cited by 10 publications

(5 citation statements)

References 20 publications

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“…Cycle threshold (Ct) and viral load correlation q-PCR results, including Ct values, were obtained from the computerized Central Virology Laboratory database. Extrapolation of the adenoviral load from Ct was calculated using known quantitated samples as a reference, a common method for quantifying viral load in clinical samples (Borg et al, 2003;Zauli et al, 2016). For adenovirus DNA copy number calibration, samples with predetermined copy numbers provided by the Quality Control for Molecular Diagnostics (QCMD) were used.…”

Section: Viral Genome Quantification By Real-time Pcr (Q-pcr)mentioning

confidence: 99%

Adenovirus load correlates with respiratory disease severity among hospitalized pediatric patients

Goikhman

Drori

Friedman

et al. 2020

International Journal of Infectious Diseases

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Objectives: Human adenoviruses (HAdVs) are common pathogens that can cause respiratory, gastrointestinal and other infections. We investigated the correlation between adenovirus viral load in clinical respiratory samples and the respiratory disease severity in pediatric patients. Methods: Medical records of patients hospitalized in the Sheba Medical Center (SMC) with confirmed adenovirus infection were retrospectively analyzed. The possible correlation between disease severity score and Real time PCR 'cycle threshold' (Ct), a proxy of viral load, was assessed in patients aged 9 years and under. In addition, Ct values of hospitalized versus community-care patient samples, positive for various respiratory viruses including adenovirus, were compared. Results: Adenovirus load in respiratory samples, as measured by Ct values, was found to be negatively correlated with respiratory disease severity in hospitalized pediatric patients aged under 9 years. Moreover, hospitalized patients presented with significantly higher Ct levels for various respiratory viruses as compared to community-care patients. Conclusion:In this study we found a correlation between Ct values obtained from adenovirus q-PCR analysis of respiratory clinical samples and disease severity in patients aged 9 years and under. Such finding may serve as a predictor of respiratory disease course in pediatric patients and will be beneficial for the differential diagnosis and treatment of pediatric patients.

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Section: Viral Genome Quantification By Real-time Pcr (Q-pcr)mentioning

confidence: 99%

Adenovirus load correlates with respiratory disease severity among hospitalized pediatric patients

Goikhman

Drori

Friedman

et al. 2020

International Journal of Infectious Diseases

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“…Quantitative PCR (qPCR)-based assays have routinely been used for the detection of hepatitis B virus and hepatitis C virus [41]. There is no report of any quantification assay for detection of any PVT1-derived transcripts.…”

Section: Discussionmentioning

confidence: 99%

Copy number-based quantification assay for non-invasive detection of PVT1-derived transcripts

Pal

Ogunwobi

2019

PLoS ONE

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BackgroundOne of the most important susceptibility loci for cancer is the 8q24 human chromosomal region. The non-protein coding gene locus plasmacytoma variant translocation 1 (PVT1) is located at 8q24 and is dysregulated in prostate cancer. PVT1 gives rise to multiple transcripts which may have different functions. Here, we describe a real-time quantitative polymerase chain reaction (qPCR)-based assay for copy number-based quantitation of PVT1 exons 4A, 4B, and 9 to enable accurate, reproducible, and quantifiable detection.MethodsPVT1 exons 4A, 4B, and 9 were cloned into a plasmid vector to create standards for subsequent creation of linear standard curves representing a broad range of concentrations. PCR was carried out using SYBR-Green signal detection to quantify PVT1 exons 4A, 4B, and 9. The efficacy of this assay was evaluated by using it to detect these transcripts in prostate epithelial and prostate cancer cell lines, normal and cancerous human prostate tissues, human serum, mouse plasma, and urine samples.ResultsThe results indicate that the assay can be used to quantify both low and high copy numbers of PVT1-derived transcripts. This is the first report of a copy number-based quantification assay for non-invasive detection of PVT1 derived transcripts.ConclusionsThis novel assay holds promise for routine non-invasive testing in diseases where PVT1 is dysregulated.

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“…The observed false positive results, hence, higher HCV antibody seroprevalence (1.67%) by Diaspot rapid enzyme immunoassay compared to that of Biorad Genscreen HCV Ag-Ab Monolisa assay (0.33%) based on 'sandwich' ELISA technique in this study contrast the outcomes of studies by other researchers who found false negative results using Diaspot rapid one-step enzyme immunoassay [18] . Current trends in transfusion service demand that initial screening of blood and blood products be based on ELISA technique using kit that both screen for the HCV core antigen and anti-HCV antibodies that improves early detection of HCV infection during the window period and subsequent confirmation of HCV status with recombinant immunoblotting assay (RIBA) or nucleic acid testing (NAT) by real-time PCR [40][41][42] . Literatures have showed that continued development of newer testing techniques have helped to significantly improve early detection and reduce the window period from 82 days to 66 days and even lower [43] .…”

Section: Discussionmentioning

confidence: 99%

Hepatitis C Virus Diagnosis in Prospective Blood Donors: Epidemiology, Optimal Testing Approach and Treatment Cut-offs

Ifechukwudebelu¹,

Kolawole²,

Ositadima³

2017

Am. J. Biomed. Sci.

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Hepatitis C virus (HCV) infection has been noted a major public health problem with no vaccines available currently. This study aims at optimizing 'safe blood' practice by advancing HCV testing beyond Diaspot rapid enzyme immunoassay in current use in Nigerian Health Institutions. Between August, 2014 and November, 2015, a total of 300 blood donors' plasma samples were screened with both Diaspot and HCV Ag-Ab enzyme-linked immunosorbent assay techniques. HCV-RNA confirmation and quantification were performed with real-time polymerase chain reaction (PCR). Alanine aminotransferase (ALT) was performed on confirmed positive blood donor for treatment purpose. The overall gender ratio and mean age of blood donors screened for HCV were 1.5:1 and 27.67 ± 7.77 years respectively. Of the 300 blood donors screened, 5 (1.67%) and 1 (0.33%) were seropositive for HCV on the basis of Diaspot and enzyme-linked immunosorbent assay (ELISA) techniques respectively. Diagnostic odds ratio showed that ELISA is nearly 9-fold a better diagnostic tool compared to Diaspot technique. Real-time PCR assay confirmed positivity of 1 (0.33%) of the blood donor for hepatitis C. HCV-RNA viral load and plasma ALT of the lone sample were 133, 209 IU/mL and 11.1 IU/L respectively. HCV prevalence among blood donors in Ekiti state is low, 0.33%. Enzyme-linked immunosorbent assay technique should be the starting point of HCV serologic screening and a surrogate technique for real-time PCR. The development of workable algorithm to reduce risks associated with blood transfusion and enhances both blood donors' and recipients' safety is highly imperative.

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In-house quantitative real-time PCR for the diagnosis of hepatitis B virus and hepatitis C virus infections

Cited by 10 publications

References 20 publications

Adenovirus load correlates with respiratory disease severity among hospitalized pediatric patients

Adenovirus load correlates with respiratory disease severity among hospitalized pediatric patients

Copy number-based quantification assay for non-invasive detection of PVT1-derived transcripts

Hepatitis C Virus Diagnosis in Prospective Blood Donors: Epidemiology, Optimal Testing Approach and Treatment Cut-offs

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