To characterize the antigenicity of nucleocapsid proteins(NP) derived from canine coronavirus(CCo V) and canine respiratory coronavirus(CRCo V) in China, the N genes of CCo V(CCo V-BJ70) and CRCo V(CRCo V-BJ202) were ...To characterize the antigenicity of nucleocapsid proteins(NP) derived from canine coronavirus(CCo V) and canine respiratory coronavirus(CRCo V) in China, the N genes of CCo V(CCo V-BJ70) and CRCo V(CRCo V-BJ202) were cloned from swabs obtained from diseased pet dogs in Beijing and then sequenced. The recombinant NPs(r NPs) were expressed in Escherichia coli and purified by nickel-affinity column and size exclusion chromatography. Sequencing data indicated that the N genes of CCo V-BJ70 and CRCo V-BJ202 belonging to two distinctly different groups were relatively conserved within each subgroup. Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) results showed that r NPs of CCo V and CRCo V were expressed efficiently and isolated with a final purity of over 95%. Western blot analysis revealed the r NP from CRCo V could cross-react with mice antisera against human coronavirus(HCo V-229 E, NL63, OC43, HKU1), while r NP of CCo V had cross-reactivity with only anti-sera against viruses belonging to the same group(HCo V-229 E and NL63). In summary, CCo V and CRCo V r NPs were successfully expressed in E. coli and showed antigenic cross-reactivity with antisera raised against human coronaviruses. These findings indicate that further serologic studies on coronavirus infections at the animal-human interface are needed.展开更多
In 2012,a novel coronavirus,initially named as human coronavirus EMC(HCoV-EMC) but recently renamed as Middle East respiratory syndrome human coronavirus(MERS-CoV),was identified in patients who suffered severe acute ...In 2012,a novel coronavirus,initially named as human coronavirus EMC(HCoV-EMC) but recently renamed as Middle East respiratory syndrome human coronavirus(MERS-CoV),was identified in patients who suffered severe acute respiratory infection and subsequent renal failure that resulted in death.Ongoing epidemiological investigations together with retrospective studies have found 61 laboratory-confirmed cases of infection with this novel coronavirus,including 34 deaths to date.This novel coronavirus is culturable and two complete genome sequences are now available.Furthermore,molecular detection and indirect immunofluorescence assay have been developed.The present paper summarises the limited recent advances of this novel human coronavirus,including its discovery,genomic characterisation and detection.展开更多
基金supported by the Mega project for Infectious Disease Research of China (2014ZX10004001-002, 2013ZX10004601)National Basic Research Program of China (2011CB504704)
文摘To characterize the antigenicity of nucleocapsid proteins(NP) derived from canine coronavirus(CCo V) and canine respiratory coronavirus(CRCo V) in China, the N genes of CCo V(CCo V-BJ70) and CRCo V(CRCo V-BJ202) were cloned from swabs obtained from diseased pet dogs in Beijing and then sequenced. The recombinant NPs(r NPs) were expressed in Escherichia coli and purified by nickel-affinity column and size exclusion chromatography. Sequencing data indicated that the N genes of CCo V-BJ70 and CRCo V-BJ202 belonging to two distinctly different groups were relatively conserved within each subgroup. Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) results showed that r NPs of CCo V and CRCo V were expressed efficiently and isolated with a final purity of over 95%. Western blot analysis revealed the r NP from CRCo V could cross-react with mice antisera against human coronavirus(HCo V-229 E, NL63, OC43, HKU1), while r NP of CCo V had cross-reactivity with only anti-sera against viruses belonging to the same group(HCo V-229 E and NL63). In summary, CCo V and CRCo V r NPs were successfully expressed in E. coli and showed antigenic cross-reactivity with antisera raised against human coronaviruses. These findings indicate that further serologic studies on coronavirus infections at the animal-human interface are needed.
基金supported by grants from the National Basic Research Program of China (2011CB504704)the State Megaproject for Infectious Disease Research of China (2011ZX10004-001)
文摘In 2012,a novel coronavirus,initially named as human coronavirus EMC(HCoV-EMC) but recently renamed as Middle East respiratory syndrome human coronavirus(MERS-CoV),was identified in patients who suffered severe acute respiratory infection and subsequent renal failure that resulted in death.Ongoing epidemiological investigations together with retrospective studies have found 61 laboratory-confirmed cases of infection with this novel coronavirus,including 34 deaths to date.This novel coronavirus is culturable and two complete genome sequences are now available.Furthermore,molecular detection and indirect immunofluorescence assay have been developed.The present paper summarises the limited recent advances of this novel human coronavirus,including its discovery,genomic characterisation and detection.
