寡核苷酸探针在传染性支气管炎病毒诊断和毒株鉴定中的应用
Oligonucleotide probes in infectious bronchitis virus diagnosis and strain identification.
作者信息
Karaca K, Palukaitis P, Naqi S
机构信息
Department of Avian and Aquatic Animal Medicine, College of Veterinary Medicine, Cornell University, Ithaca, NY.
出版信息
J Virol Methods. 1993 May;42(2-3):293-300. doi: 10.1016/0166-0934(93)90040-x.
Genomic RNA fingerprints of infectious bronchitis virus (IBV) strains M41 and Conn46 were prepared to identify T1 RNase-resistant oligonucleotides 'unique' to each of the two IBV strains. Such oligonucleotides were subsequently eluted from the gels and their nucleotide sequences determined. When oligonucleotide probes of those sequences were synthesized and used in a dot-blot hybridization assay, the probes lacked IBV strain-specificity and reacted with the RNAs of homologous as well as heterologous IBV strains. Based on these results, the methods used in this study need to be applied to a large number of oligonucleotide probes, to find one or a few that might be suitable as IBV strain- or serotype-specific oligonucleotide probes.
制备了传染性支气管炎病毒（IBV）毒株M41和Conn46的基因组RNA指纹图谱，以鉴定这两种IBV毒株各自“独特”的抗T1核糖核酸酶寡核苷酸。随后从凝胶中洗脱这些寡核苷酸并测定其核苷酸序列。当合成这些序列的寡核苷酸探针并用于斑点印迹杂交试验时，这些探针缺乏IBV毒株特异性，能与同源及异源IBV毒株的RNA发生反应。基于这些结果，本研究中使用的方法需要应用于大量的寡核苷酸探针，以找到一个或几个可能适合作为IBV毒株或血清型特异性寡核苷酸探针的探针。
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