2023
DOI: 10.3389/fvets.2022.1059934
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Development of a multienzyme isothermal rapid amplification and lateral flow dipstick combination assay for bovine coronavirus detection

Abstract: Bovine coronavirus (BCoV) is a major cause of infectious disease in cattle, causing huge economic losses to the beef and dairy industries worldwide. BCoV can infect humans and multiple other species of animals. A rapid, reliable, and simple test is needed to detect BCoV infection in suspected farms. In this study, we developed a novel multienzyme isothermal rapid amplification (MIRA) and lateral flow dipstick (LFD) combination assay, targeting a highly conserved region of the viral nucleocapsid (N) gene for BC… Show more

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Cited by 10 publications
(13 citation statements)
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“…However, only 16.6% of samples from A State tested positive for the tlh gene using the PCR assay, and all samples from the other three States tested negative. Our results suggest that the MIRA-LFD assay is as sensitive as the qPCR assay, taken together with previous studies [31,38,41]. Interestingly, all oyster samples from all four States tested negative for the tdh and trh genes using the MIRA-LFD, PCR, and qPCR assays.…”
Section: Detection Of Tlh Trh and Tdh Genes From Oyster Samplessupporting
confidence: 80%
“…However, only 16.6% of samples from A State tested positive for the tlh gene using the PCR assay, and all samples from the other three States tested negative. Our results suggest that the MIRA-LFD assay is as sensitive as the qPCR assay, taken together with previous studies [31,38,41]. Interestingly, all oyster samples from all four States tested negative for the tdh and trh genes using the MIRA-LFD, PCR, and qPCR assays.…”
Section: Detection Of Tlh Trh and Tdh Genes From Oyster Samplessupporting
confidence: 80%
“…Interestingly, CkChpV alone can be precisely detected, with no cross-reaction with other common viruses, thereby allowing the accurate interpretation of data. MIRA combined with LFD can completely replace conventional PCR and only requires a constant temperature instrument in areas with poor research conditions for food safety, pathogen detection, and other application related to nucleic acid testing ( Ji et al, 2022 ).…”
Section: Discussionmentioning
confidence: 99%
“…This event occurs at both the forward and reverse end of the DNA. Once the primer recombinase complex binds to the ssDNA, DNA polymerase synthesizes the new complementary DNA strand [ 69 ]. The use of helicase (gp41) and recombinases ( Streptomyces coelicolor recA, SC-recA) allows for rapid amplification that can happen between 10–30 min at a constant temperature between 35–42 °C [ 68 , 69 ].…”
Section: Raa and Mira With Paper Microfluidicsmentioning
confidence: 99%
“…Once the primer recombinase complex binds to the ssDNA, DNA polymerase synthesizes the new complementary DNA strand [ 69 ]. The use of helicase (gp41) and recombinases ( Streptomyces coelicolor recA, SC-recA) allows for rapid amplification that can happen between 10–30 min at a constant temperature between 35–42 °C [ 68 , 69 ]. MIRA has also been demonstrated with paper strips and paper microfluidic platforms [ 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 ].…”
Section: Raa and Mira With Paper Microfluidicsmentioning
confidence: 99%
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