Peer-reviewed veterinary case report
Development of a loop-mediated isothermal amplification assay for rapid detection of bovine parvovirus.
- Journal:
- Journal of virological methods
- Year:
- 2013
- Authors:
- Luo, Ji-guan et al.
- Affiliation:
- College of Veterinary Medicine · China
Plain-English summary
Researchers have created a new test called a loop-mediated isothermal amplification (LAMP) assay to quickly detect bovine parvovirus (BPV), a virus that can affect cattle. This test uses specific primers to target parts of the virus's DNA and can identify very small amounts of the virus, making it much more sensitive than traditional testing methods. The LAMP assay can be done in about 45 minutes and is easy to use, whether in a lab or out in the field. When tested on samples from cattle, the results matched those from the standard testing method, showing that this new approach is both effective and reliable. Overall, the LAMP assay is a promising tool for quickly diagnosing BPV infections in cattle.
Abstract
A loop-mediated isothermal amplification (LAMP) assay was developed for detection of bovine parvovirus (BPV) DNA. Four primers were designed to recognize six distinct regions on the target DNA based on a highly conserved sequence in the VP2 region of the BPV genome. The optimized LAMP reaction conditions were 8 mM Mg²⁺, 1.2 mM betaine, and an incubation at 63°C for 45 min. After amplification the products were detected either by observing a ladder pattern following gel electrophoresis, observation of turbidity, or a color change with the addition of SYBR Green I to the reaction tube. The detection limit of the LAMP assay was 9 copies of BPV-DNA and was 100 times more sensitive than conventional PCR. A ladder pattern of bands after gel electrophoresis was observed for only BPV isolates and showed that the BPV LAMP assay was highly specific without any cross-reactivity with other related viruses. The LAMP assay was evaluated further using 59 field samples and the results were comparable to conventional PCR. The LAMP assay is a simple, rapid and economic detection method; it can provide a useful technique suitable for detection of BPV infection in both field conditions and laboratory settings.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/22584269/