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IBDV强、弱毒株感染SPF鸡后其病毒基因拷贝数在不同组织内的动态分析研究
郑丽
0
(中国兽医药品监察所)
摘要:
为研究IBDV强毒和弱毒在感染鸡体内的动态分布情况,以探究 IBDV不同毒株对鸡的致病情况,。本试验实验根据IBDV 弱毒株B87 E6和强毒株 BC6/85的基因序列,合成non-vvIBDV 的TaqMan-MGB通用探针PN及其相关引物,建立了基于该探针的实时荧光定量RT-PCR方法,并构建标准质粒绘制了标准曲线,进行了特异性、灵敏度等检测。结果显示:所设引物扩增的片段与预期目的片段大小相符,标准曲线有良好的线性关系,特异性、灵敏度均良好。根据已建立的上述方法,对弱毒B87 E6和强毒BC6/85感染SPF鸡后病毒在鸡体内的组织分布进行检测和初步研究。结果表明,感染后第3d,试验实验组1(B87 E6单独感染组)在第3d的法氏囊病毒基因组拷贝数低于试验实验组2(BC6/85单独感染组)和试验实验组3(B87与BC6/85共感染组)。试验实验组3法氏囊中IBDV基因组拷贝数总体并不比试验实验组1、试验实验组2的大,可能是推测同时接种IBDV强、弱毒时,强毒BC6/85对法氏囊组织的感染起主要作用。此外,病毒的主要复制位点仍然是法氏囊组织。
关键词:  IBDV  荧光定量RT-PCR  动态分布
DOI:
投稿时间:2017-05-12修订日期:2017-06-01
基金项目:国家微生物资源平台(NIMR-5)
Dynamic Analysis of Viral Gene Copies in Different Tissues of SPF Chickens Infected with IBDV Virulent and Attenuated Stains
(China Institute of Veterinary Drug Control)
Abstract:
In order to study the dynamic distribution of IBDV virulent and attenuated IBDV Stains chickens in infected chickens, and the pathogenicity of IBDV two strains was studied. According to the genes of IBDV attenuated strain B87 E6 and virulent strain BC6 / 85, and the TaqMan-MGB universal probe PN of the non-vvIBDV of a published paper was synthesized. And the real-time quantitative RT-PCR method based on this probe PN method was established .The standard curve was drawn, the method was tested for specificity, sensitivity and so on. The results showed that the primers were in good agreement with the expected size of the target, and the correlation coefficient, specificity and sensitivity of probe PN is good. Based on the method had established, the tissue distribution of attenuated B87 E6 and virulent BC6 / 85 infected SPF chicken infected with attenuated strain B87 E6 and virulent strain BC6 / 85virus wereas detected and studied. The results showed that, at the third day ,the gene copy copies number of G1 bursa genome was significantly lower in BC7 / 85 infection group and B87 and BC6 / 85 co-infected group at the third day of B87 infected groupthan G2 and G3. There was no significant difference in copy number of viral genes in each group at other time points. In the test group 3, the copy number of IBDV genome in of the bursa was not significantly higher than that in the group 1 and the group 2, which was probably the virulent BC6 / 85 had the main effect on the infection of bursa when there had the two strain. In addition, the main site of IBDV replication is the bursa.
Key words:  IBDV, Fluorescence quantitative RT – PCR , Dynamic distribution

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