Kwon H M, Saif L J, Jackwood D J
Department of Veterinary Preventive Medicine, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster 44691, USA.
J Vet Med Sci. 1998 May;60(5):589-97. doi: 10.1292/jvms.60.589.
The diversity in selected regions of the transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV) genomes was analyzed among known TGEV and PRCV strains and field isolates. The N-terminal half of the spike (S) glycoprotein gene and open reading frames (ORF) 3, 3-1 and 4 were amplified by reverse transcriptase reaction and polymerase chain reaction (RT/PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. Reference TGEV strains (Miller and Purdue) and a PRCV strain (ISU-1), and TGEV and PRCV field isolates were analyzed. Based on the size of the ORF 3, 3-1 and 4 RT/PCR products, TGEV and PRCV strains could be quickly and easily differentiated into three groups designated TGEV Miller, Purdue types and PRCV. By RFLP analysis of the N-terminal region of the S glycoprotein gene and ORFs 3, 3-1 and 4, TGEV and PRCV strains were differentiated into five groups using the restriction enzyme Sau3AI. Sequence analysis of a PCR product in the ORFs 3, 3-1 and 4 from virulent and attenuated Miller strains demonstrated additional differences in that region which have been correlated with a change in virulence of TGEV isolates.
在已知的传染性胃肠炎病毒（TGEV）和猪呼吸道冠状病毒（PRCV）毒株及田间分离株中，分析了TGEV和PRCV基因组选定区域的多样性。通过逆转录反应和聚合酶链反应（RT/PCR）扩增刺突（S）糖蛋白基因的N端一半以及开放阅读框（ORF）3、3-1和4，并使用扩增DNA的限制性片段长度多态性（RFLP）模式进行分析。对参考TGEV毒株（Miller和Purdue）、一株PRCV毒株（ISU-1）以及TGEV和PRCV田间分离株进行了分析。根据ORF 3、3-1和4 RT/PCR产物的大小，TGEV和PRCV毒株可快速、轻松地分为三组，分别命名为TGEV Miller、Purdue型和PRCV。通过对S糖蛋白基因N端区域以及ORF 3、3-1和4进行RFLP分析，使用限制性内切酶Sau3AI将TGEV和PRCV毒株分为五组。对强毒和弱毒Miller毒株ORF 3、3-1和4中的PCR产物进行序列分析，结果表明该区域存在其他差异，这些差异与TGEV分离株毒力的变化有关。
