Abstract
We applied a probe-based real-time loop-mediated isothermal amplification (Cy5-RTqLAMP) technique targeting the avian reovirus (ARV) S3 gene to develop a rapid, sensitive, and specific method for virus detection and quantification. This test specifically detected the presence of ARV, but not other viruses or bacteria present in clinical or artificially spiked samples, including Newcastle disease virus, infectious bursal disease virus, fowl adenovirus, Marek’s disease virus, Escherichia coli, and Salmonella spp. This test can detect ARV in less than one hour with an analytical sensitivity of 10 viral gene copies and 1 fg of total cDNA. The Cy5-RTqLAMP does not yield false positive results and is 100 times more sensitive than conventional PCR. This test was shown to be able to detect the presence of ARV in clinical samples. A similar strategy may be used for detection of other important human and animal viral pathogens.
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Acknowledgements
These studies were funded by Grant BT/PR6232/GBD/27/392/2012, provided by the Department of Biotechnology, Government of India. The authors are also thankful to the Director of IVRI for providing necessary research facilities.
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Kumar, D., Chauhan, T.K.S., Agarwal, R.K. et al. A double-stranded probe coupled with isothermal amplification for qualitative and quantitative detection of avian reovirus. Arch Virol 162, 979–985 (2017). https://doi.org/10.1007/s00705-016-3184-1
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DOI: https://doi.org/10.1007/s00705-016-3184-1