Human metapneumovirus(hMPV) is a recently identified respiratory virus more like human respiratory syncytial virus in clinical symptoms.Matrix protein(M) is one of the most important structural proteins.For further st...Human metapneumovirus(hMPV) is a recently identified respiratory virus more like human respiratory syncytial virus in clinical symptoms.Matrix protein(M) is one of the most important structural proteins.For further studying of hMPV,the full length of M genes from the recombinant plasmid pUCm-M1816 and pUCm-M1817 were cloned by PCR and sub-cloned into the pET30a(+) vector,which is a prokaryotic expression vector,after dual-enzyme digestion with Bam HI and Xho I.The positive recombinated plasmids were transformed into E.coli BL21(DE3) and expressed under the inducing of IPTG.Target proteins were characterized by SDSPAGE and Western blotting.In this article,we’ve successfully constructed the recombinated plasmids pET30a-M1816 and pET30a-M1817 which have correct open reading frames confirmed by dual-enzyme digestion analysis and sequencing. The fusion proteins with 6·His-N were highly produced after inducing by 1mmol/L IPTG at 37℃.A unique protein band with approximate 27.6 kD was characterized by SDS-PAGE.Most of the target protein existed in inclusion body.Western blot analysis showed that the target protein has specific binding reaction to rabbit antiserum against polypeptides of the matrix protein of hMPV.So the M genes were highly expressed in the prokaryotic system and the expressed M proteins have specific antigenic activities.It can be used for further studying of hMPV infections in Beijing.展开更多
快速荧光灶抑制试验(RFFIT)是世界卫生组织(World Health Organisation,WHO)推荐的狂犬病病毒中和抗体检测标准方法之一,国内进行RFFIT检测的不同实验室之间互评可促进该方法推广时的标准化。病毒病预防控制所和中国药品生物制品检定所...快速荧光灶抑制试验(RFFIT)是世界卫生组织(World Health Organisation,WHO)推荐的狂犬病病毒中和抗体检测标准方法之一,国内进行RFFIT检测的不同实验室之间互评可促进该方法推广时的标准化。病毒病预防控制所和中国药品生物制品检定所对200例人血清进行了双重平行RFFIT检测。结果显示双方检测中和抗体效价几何均值(genomic means titration,GMT)为1.89、1.84I U/mL,差别无统计学意义(t=-0.94,P>0.05);血清中和抗体阳性率为95.00%、93.00%,差别无统计学意义(χ2=0.71,P>0.05);双方结果定性一致率为89.00%,发生结果定性不一致的22例(11.00%)血清双方检测结果均<1I U/mL;67例双方检测结果均<1I U/mL的血清样品中有22例(32.84%)发生结果定性不一致。展开更多
文摘Human metapneumovirus(hMPV) is a recently identified respiratory virus more like human respiratory syncytial virus in clinical symptoms.Matrix protein(M) is one of the most important structural proteins.For further studying of hMPV,the full length of M genes from the recombinant plasmid pUCm-M1816 and pUCm-M1817 were cloned by PCR and sub-cloned into the pET30a(+) vector,which is a prokaryotic expression vector,after dual-enzyme digestion with Bam HI and Xho I.The positive recombinated plasmids were transformed into E.coli BL21(DE3) and expressed under the inducing of IPTG.Target proteins were characterized by SDSPAGE and Western blotting.In this article,we’ve successfully constructed the recombinated plasmids pET30a-M1816 and pET30a-M1817 which have correct open reading frames confirmed by dual-enzyme digestion analysis and sequencing. The fusion proteins with 6·His-N were highly produced after inducing by 1mmol/L IPTG at 37℃.A unique protein band with approximate 27.6 kD was characterized by SDS-PAGE.Most of the target protein existed in inclusion body.Western blot analysis showed that the target protein has specific binding reaction to rabbit antiserum against polypeptides of the matrix protein of hMPV.So the M genes were highly expressed in the prokaryotic system and the expressed M proteins have specific antigenic activities.It can be used for further studying of hMPV infections in Beijing.
文摘快速荧光灶抑制试验(RFFIT)是世界卫生组织(World Health Organisation,WHO)推荐的狂犬病病毒中和抗体检测标准方法之一,国内进行RFFIT检测的不同实验室之间互评可促进该方法推广时的标准化。病毒病预防控制所和中国药品生物制品检定所对200例人血清进行了双重平行RFFIT检测。结果显示双方检测中和抗体效价几何均值(genomic means titration,GMT)为1.89、1.84I U/mL,差别无统计学意义(t=-0.94,P>0.05);血清中和抗体阳性率为95.00%、93.00%,差别无统计学意义(χ2=0.71,P>0.05);双方结果定性一致率为89.00%,发生结果定性不一致的22例(11.00%)血清双方检测结果均<1I U/mL;67例双方检测结果均<1I U/mL的血清样品中有22例(32.84%)发生结果定性不一致。
