新城疫病毒融合蛋白裂解位点基因光敏生物素探针的制备及初步应用

The Preparation and Preliminary Application of the Photobiotin Probe of the Fusion Cleavage Site Gene of Newcastle Disease Virus (NDV)

将重组质粒ｐＵＣＬａＦｃ和ｐＵＣＦ４６Ｆｃ用ＥｃｏＲＩ和ＳａｌⅠ酶切，回收新城疫病毒融合蛋白裂解位点（Ｆｃ）基因，将此Ｆｃ片段用光敏生物素标记，制备出Ｆｃ探针。该探针能够与新城疫病毒（ＮＤＶ）的强毒株Ｆ４６Ｅ９、中毒株Ｍ株和弱毒株ＬａＳｏｔａＥ４株杂交，而不与对照的传染性支气管炎病毒（ＩＢＶ）、减蛋综合征（ＥＤＳ－７６）病毒杂交，说明探针是特异的。用ｐＵＣＬａＦｃ的Ｆｃ探针检测了各５份ＮＤＶ强中弱毒株的尿囊液，均为阳性。但强毒株和弱毒株的Ｆｃ探针都能与所用的强、中、弱毒株杂交，说明该Ｆｃ探针尚不能区分强、弱毒株。

The recombinant plasmid pUCLaFc and pUCF46Fc had been digested, and the Fc DNA fragments recovered, using which the photobiotin probes had been prepared. The probe could hybrid with the RNA of virulent strain F46E9, the midvirulent M and the avirulent LaSotaE4 strains, while not hybrid with other RNA controls of infectious bronchitis virus (IBV) and egg drop syndrome 76 virus (EDS 76 virus), suggesting that the probes might be specific to the NDV. 5 allantoic fluid specimens of F46E9、M and LaSotaE4 each have been detected respectively, using pUCLaFc Fc probe getting the positive results. It was proved that the probes could react with virulent, midvirulent and avirulent strains of NDV, indicating the probes not be able to differentiate the virulence of different NDV strains.