Vieler E, Schlapp T, Anders C, Herbst W
Institut für Hygiene und Infektionskrankheiten, Tiere der Justus-Liebig-Universität Giessen, Federal Republic of Germany.
Arch Virol. 1995;140(7):1215-23. doi: 10.1007/BF01322747.
The genomic relationship of porcine hemagglutinating encephalomyelitis virus (HEV) to bovine coronavirus (BCV) and human coronavirus (HCV) strain OC43 was examined by dot blot hybridization assays. Two BCV S gene-specific probes were generated by polymerase chain reaction from the avirulent L9-strain of BCV. Probes were located in the S1 and the S2 region of the peplomeric (S) glycoprotein gene. The S1 probe (726 bp) hybridized with BCV and HCV-OC43, but not with HEV under moderate stringency hybridization conditions (50 degrees C). Only slight signals were present with mouse hepatitis virus (MHV) and no signals were observed with feline infectious peritonitis virus (FIPV) or canine coronavirus (CCV). At high stringency conditions (60 degrees C) the S1 probe hybridized with BCV only. Using the S2 probe (680 bp) under moderate stringency conditions, hybridization signals were obtained with BCV, HCV-OC43 and HEV (strains 67N, NT9, VW572). The signals obtained by the three HEV strains were altogether weaker than with BCV and HCV-OC43. The S2 probe did not react with MHV, FIPV and CCV. At high stringency the S2-specific probe hybridized with BCV and HCV-OC43 but did not hybridize with HEV. Nucleotide sequence analysis of the region covering the S2 probe in HEV revealed 92.6% nucleotide sequence homology to BCV and 91.9% to HCV-OC43. In contrast, the region covering the S1 probe in HEV could not be amplified using the BCV S1-specific primers. The hybridization and sequencing results thus indicate a closer genomic relationship between BCV and HCV-OC43 than there is between HEV and BCV or HCV-OC43 respectively.
通过斑点印迹杂交试验检测了猪血凝性脑脊髓炎病毒(HEV)与牛冠状病毒(BCV)以及人冠状病毒(HCV)OC43株之间的基因组关系。通过聚合酶链反应从BCV无毒L9株中产生了两个BCV S基因特异性探针。探针位于纤突(S)糖蛋白基因的S1和S2区域。在中等严格杂交条件(50℃)下,S1探针(726 bp)与BCV和HCV-OC43杂交,但不与HEV杂交。小鼠肝炎病毒(MHV)仅有微弱信号,而猫传染性腹膜炎病毒(FIPV)或犬冠状病毒(CCV)未观察到信号。在高严格条件(60℃)下,S1探针仅与BCV杂交。在中等严格条件下使用S2探针(680 bp),可获得与BCV、HCV-OC43和HEV(67N、NT9、VW572株)的杂交信号。三种HEV株获得的信号总体上比BCV和HCV-OC43的信号弱。S2探针不与MHV、FIPV和CCV反应。在高严格条件下,S2特异性探针与BCV和HCV-OC43杂交,但不与HEV杂交。对HEV中覆盖S2探针的区域进行核苷酸序列分析发现,其与BCV的核苷酸序列同源性为92.6%,与HCV-OC43的同源性为91.9%。相比之下,使用BCV S1特异性引物无法扩增HEV中覆盖S1探针的区域。因此,杂交和测序结果表明,BCV与HCV-OC43之间的基因组关系比HEV与BCV或HCV-OC43之间的关系更密切。