ArticleBMC infectious diseases2002
Development of real-time NASBA assays with molecular beacon detection to quantify mRNA coding for HHV-8 lytic and latent genes.
Article in BMC infectious diseases, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 51 citations in OpenAlex.
- RPA-CRISPR/Cas12a mediated isothermal amplification for visual detection ofFrontiers in cellular and infection microbiology · 2023Article
- Three-Way Junction-Induced Isothermal Amplification with High Signal-to-Background Ratio for Detection of Pathogenic Bacteria.Sensors (Basel, Switzerland) · 2021Article
- Development of a real-time nucleic acid sequence-based amplification assay for the rapid detection of Salmonella spp. from food.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2019Article
- HCV Detection, Discrimination, and Genotyping Technologies.Sensors (Basel, Switzerland) · 2018Review
- Molecular beacons: a novel optical diagnostic tool.Archivum immunologiae et therapiae experimentalis · 2013Review
- Application of quantitative PCR for the detection of microorganisms in water.Analytical and bioanalytical chemistry · 2013Review
- Molecular beacon probes-base multiplex NASBA Real-time for detection of HIV-1 and HCV.Iranian journal of microbiology · 2012Article
- Human herpesvirus 8--a novel human pathogen.Virology journal · 2005Review
- Human herpesvirus 8 load in matched serum and plasma samples of patients with AIDS-associated Kaposi's sarcoma.Journal of clinical microbiology · 2003Article
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundHuman herpesvirus-8 (HHV-8) is linked to the pathogenesis of Kaposi's sarcoma (KS), and the HHV-8 DNA load in peripheral blood mononuclear cells (PBMC) is associated with the clinical stage of KS. To examine the expression of HHV-8 in PBMC, four HHV-8 mRNA specific NASBA assays were developed
methodsWe have developed four quantitative nucleic acid sequence-based amplification assays (NASBA-QT) specifically to detect mRNA coding for ORF 73 (latency-associated nuclear antigen, LANA), vGCR (a membrane receptor), vBcl-2 (a viral inhibitor of apoptosis) and vIL-6 (a viral growth factor). The NASBA technique amplifies nucleic acids without thermocycling and mRNA can be amplified in a dsDNA background. A molecular beacon is used during amplification to enable real-time detection of the product. The assays were tested on PBMC samples of two AIDS-KS patients from the Amsterdam Cohort.
resultsFor all four assays, the limit of detection (LOD) of 50 molecules and the limit of quantification (LOQ) of 100 molecules were determined using in vitro transcribed RNA. The linear dynamic range was 50 to 10(7) molecules of HHV-8 mRNA. We found HHV-8 mRNA expression in 9 out of the 10 tested samples.
conclusionThese real-time NASBA assays with beacon detection provide tools for further study of HHV-8 expression in patient material.
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