1. Simultaneous and visual detection of infectious bronchitis virus and Newcastle disease virus by multiple LAMP and lateral flow dipstick.
- Author
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Wu X, Song Z, Zhai X, Zuo L, Mei X, Xiang R, Kang Z, Zhou L, and Wang H
- Subjects
- Animals, China, Chromatography methods, Coronavirus Infections diagnosis, Coronavirus Infections virology, Infectious bronchitis virus isolation & purification, Newcastle Disease virology, Newcastle disease virus isolation & purification, Nucleic Acid Amplification Techniques methods, Poultry Diseases virology, Chickens, Chromatography veterinary, Coronavirus Infections veterinary, Newcastle Disease diagnosis, Nucleic Acid Amplification Techniques veterinary, Poultry Diseases diagnosis
- Abstract
Infectious bronchitis virus (IBV) and Newcastle disease virus (NDV) are both important viruses seriously affecting poultry industry worldwide. In this study, reverse-transcription LAMP (RT-LAMP) was combined with lateral flow dipstick (LFD) forming a novel detection tool which could simultaneously detect IBV and NDV visually. Primers targeted the 5'-untranslated region (5'-UTR) of IBV genome and the conserved region of NDV large polymerase gene (LP). The specificity and sensitivity of this multiple reverse transcription-LAMP-LFD (mRT-LAMP-LFD) assay were compared with those of conventional RT-PCR, nested RT-PCR (nRT-PCR), quantification RT-PCR (qRT-PCR), and RT-LAMP monitored by electrophoresis. No non-specific amplifications were observed when the assays were tested with unrelated viruses. According to the sensitivity study, when detecting IBV or NDV alone, the lowest detection limits of mRT-LAMP-LFD were 100.8 IBV RNA copies/reaction and 100.7 NDV RNA copies/reaction. Furthermore, when detecting IBV and NDV simultaneously, the lowest detection limit was the same as that of the single detection assays. In the clinical sample study, mRT-LAMP-LFD performed the best among these assays. When tested with IBV or NDV single infected samples, the mean detection rates were 98.65% and 97.25%, respectively. In the IBV and NDV co-infected sample study, the mean detection rates of IBV and NDV were both 95%. This study showed that mRT-LAMP-LFD was a promising qualitative detection tool suitable for field single or multiple IBV and NDV detection., (© 2019 Poultry Science Association Inc.)
- Published
- 2019
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