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Multiplex detection of six swine viruses on an integrated centrifugal disk using loop-mediated isothermal amplification.
- Source :
- Journal of Veterinary Diagnostic Investigation; May2019, Vol. 31 Issue 3, p415-425, 11p
- Publication Year :
- 2019
-
Abstract
- Advances in molecular testing and microfluidic technologies have opened new avenues for rapid detection of animal viruses. We used a centrifugal microfluidic disk (CMFD) to detect 6 important swine viruses, including foot-and-mouth disease virus, classical swine fever virus, porcine reproductive and respiratory swine virus–North American genotype, porcine circovirus 2, pseudorabies virus, and porcine parvovirus. Through integrating the loop-mediated isothermal amplification (LAMP) method and microfluidic chip technology, the CMFD could be successfully performed at 62℃ in 60 min. The detection limit of the CMFD was 3.2 × 10<superscript>2</superscript> copies per reaction, close to the sensitivity of tube-type LAMP turbidity methods (1 × 10<superscript>2</superscript> copies per reaction). In addition, the CMFD was highly specific in detecting the targeted viruses with no cross-reaction with other viruses, including porcine epidemic diarrhea virus, transmissible gastroenteritis virus, and porcine rotavirus. The coincidence rate of CMFD and conventional PCR was ~94%; the CMFD was more sensitive than conventional PCR for detecting mixed viral infections. The positive detection rate of 6 viruses in clinical samples by CMFD was 44.0% (102 of 232), whereas PCR was 40.1% (93 of 232). Thirty-six clinical samples were determined to be coinfected with 2 or more viruses. CMFD can be used for rapid, sensitive, and accurate detection of 6 swine viruses, offering a reliable assay for monitoring these pathogens, especially for detecting viruses in widespread mixed-infection clinical samples. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 10406387
- Volume :
- 31
- Issue :
- 3
- Database :
- Complementary Index
- Journal :
- Journal of Veterinary Diagnostic Investigation
- Publication Type :
- Academic Journal
- Accession number :
- 136889529
- Full Text :
- https://doi.org/10.1177/1040638719841096