Peer-reviewed veterinary case report
Development of a loop-mediated isothermal amplification method for rapid detection of porcine boca-like virus.
- Journal:
- Journal of virological methods
- Year:
- 2012
- Authors:
- Li, Bin et al.
- Affiliation:
- Institute of Veterinary Medicine · China
Plain-English summary
Researchers have developed a new test to quickly and accurately detect a virus called porcine boca-like virus (Pbo-likeV) in pigs, particularly those suffering from a condition known as post-weaning multisystemic wasting syndrome (PMWS). This test, called loop-mediated isothermal amplification (LAMP), can identify the virus using a simple method that doesn't require special equipment. It works by targeting specific parts of the virus's genetic material and can detect very small amounts of the virus, making it much more sensitive than traditional testing methods. Importantly, this new test does not confuse Pbo-likeV with other common pig viruses, ensuring accurate results. Overall, the LAMP test is a promising tool for detecting this virus in both labs and on farms.
Abstract
The porcine boca-like virus (Pbo-likeV) was recently discovered in Swedish pigs with post-weaning multisystemic wasting syndrome (PMWS). In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for rapid, specific and sensitive detection of Pbo-likeV. A set of four primers specific for six regions of Pbo-likeV VP1/2 genes was designed with the online software. The reaction temperature and time were optimized to 65 °C and 60 min, respectively. LAMP products were detected by agarose gel electrophoresis or by visual inspection of a color change due to addition of fluorescent dye. The developed method was highly specific for detection of Pbo-likeV, and no cross-reaction was observed with other swine viruses, such as porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and classic swine fever virus (CSFV) found commonly in China. The lower detection limit of the LAMP assay was approximately 10 copies per reaction, and it was 100 times more sensitive than that of conventional PCR. Furthermore, the efficiency of LAMP for detection Pbo-likeV in clinical samples was comparable to PCR and sequencing. These results showed that the LAMP assay is a simple, rapid, sensitive and specific technique for detection of Pbo-likeV, and the procedure of LAMP does not rely on any special equipment. It has capacity for the detection of Pbo-likeV both in the laboratory and on farms.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/22172971/