ArticleVirology journal2011
Development of one-step SYBR Green real-time RT-PCR for quantifying bovine viral diarrhea virus type-1 and its comparison with conventional RT-PCR.
Article in Virology journal, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 27 citations in OpenAlex.
- Article
- Diagnosis of bovine viral diarrhea virus: an overview of currently available methods.Frontiers in microbiology · 2024Review
- Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip.Animals : an open access journal from MDPI · 2021Article
- Article
- Two stage, nested isothermal amplification in a single tube.The Analyst · 2021Article
- Upregulation of interferon-alpha gene in bovine embryos produced in vitro in response to experimental infection with noncytophatic bovine-viral-diarrhea virus.Molecular biology reports · 2020Article
- Article
- Detection of bovine viral diarrhea virus genotype 1 in aerosol by a real time RT-PCR assay.BMC veterinary research · 2020Article
- Multiplex detection of six swine viruses on an integrated centrifugal disk using loop-mediated isothermal amplification.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2019Article
- A newly developed BVDV-1 RT-qPCR Taqman assay based on Italian isolates: evaluation as a diagnostic tool.Folia microbiologica · 2017Article
- Mechanism of curcumin resistance to human cytomegalovirus in HELF cells.BMC complementary and alternative medicine · 2014Article
- Article
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Authors and funding
9 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundBovine viral diarrhea virus (BVDV) is a worldwide pathogen in cattle and acts as a surrogate model for hepatitis C virus (HCV). One-step real-time fluorogenic quantitative reverse transcription polymerase chain reaction (RT-PCR) assay based on SYBR Green I dye has not been established for BVDV detection. This study aims to develop a quantitative one-step RT-PCR assay to detect BVDV type-1 in cell culture.
resultsOne-step quantitative SYBR Green I RT-PCR was developed by amplifying cDNA template from viral RNA and using in vitro transcribed BVDV RNA to establish a standard curve. The assay had a detection limit as low as 100 copies/ml of BVDV RNA, a reaction efficiency of 103.2%, a correlation coefficient (R2) of 0.995, and a maximum intra-assay CV of 2.63%. It was 10-fold more sensitive than conventional RT-PCR and can quantitatively detect BVDV RNA levels from 10-fold serial dilutions of titrated viruses containing a titer from 10(-1) to 10(-5) TCID50, without non-specific amplification. Melting curve analysis showed no primer-dimers and non-specific products.
conclusionsThe one-step SYBR Green I RT-PCR is specific, sensitive and reproducible for the quantification of BVDV in cell culture. This one-step SYBR Green I RT-PCR strategy may be further optimized as a reliable assay for diagnosing and monitoring BVDV infection in animals. It may also be applied to evaluate candidate agents against HCV using BVDV cell culture model.
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