Verbeek A, Dea S, Tijssen P
Centre de Recherche en Virologie, Institut Armand-Frappier, Université du Québec, Laval-des-Rapides, Canada.
Arch Virol. 1991;121(1-4):199-211. doi: 10.1007/BF01316754.
Genomic relationships between turkey and bovine coronavirus (TCV and BCV), which are currently placed in distinct antigenic groups, were demonstrated by hybridization using specific cDNA probes. BCV-specific recombinant plasmid probes p 52, p 27, and p 247, holding inserts derived from (probably nonstructural) genes, and plasmids pN 17 and pN 9 holding the N and M gene, respectively, permitted the detection of isolates of both BCV and TCV with similar sensitivities. Similarly, probing supernatants of cell cultures infected with several isolates of TCV, using probes pN 17 and pM 78, respectively holding the N gene of BCV and TCV, resulted in equally intense detection signals. Only a slight detection of MHV-3, which is antigenically related to BCV, was observed, whereas the probes did not allow the detection of IBV, TGEV, and HCV-229E, which are placed in antigenic groups separate from those of BCV and TCV. Detection of TCV was improved by hybridization with BCV-specific single-stranded (ss) probes holding sequences of the N and M genes and synthesized by the polymerase chain reaction. Diagnosis of TCV in 134 clinical samples by hybridization was better with PCR-produced ss BCV-specific probes than with ds PCR-produced probes or a combination of six recombinant plasmid probes holding non-overlapping BCV-specific cDNA sequences. Detection signals were absent when probing clinical samples with 32P-labelled pUC-DNA.
目前分属于不同抗原组的火鸡冠状病毒(TCV)和牛冠状病毒(BCV)之间的基因组关系,通过使用特异性cDNA探针进行杂交得以证实。BCV特异性重组质粒探针p 52、p 27和p 247,其插入片段源自(可能是非结构)基因,以及分别含有N基因和M基因的质粒pN 17和pN 9,能够以相似的灵敏度检测BCV和TCV的分离株。同样地,分别使用含有BCV和TCV的N基因的探针pN 17和pM 78,对感染多种TCV分离株的细胞培养上清液进行检测,得到的检测信号强度相同。仅观察到与BCV有抗原相关性的MHV - 3有轻微检测信号,而这些探针无法检测出IBV、TGEV和HCV - 229E,它们分属于与BCV和TCV不同的抗原组。通过与由聚合酶链反应合成的、含有N基因和M基因序列的BCV特异性单链(ss)探针杂交,提高了TCV的检测效果。用PCR制备的ss BCV特异性探针通过杂交诊断134份临床样本中的TCV,比用ds PCR制备的探针或六种含有不重叠BCV特异性cDNA序列的重组质粒探针组合效果更好。用32P标记的pUC - DNA探测临床样本时未出现检测信号。