Field isolates of transmissible gastroenteritis virus differ at the molecular level from the Miller and Purdue virulent and attenuated strains and from porcine respiratory coronaviruses
Field isolates of transmissible gastroenteritis virus differ at the molecular level from the Miller and Purdue virulent and attenuated strains and from porcine respiratory coronaviruses
复制标题
DOI：
10.1292/jvms.60.589
复制
发表时间：
1998-05-01
影响因子：
1.2
通讯作者：
Jackwood, DJ
中科院分区：
文献类型：
--
作者：
Kwon, HM;Saif, LJ;Jackwood, DJ
在已知的 TGEV 和 PRCV 毒株和野外分离株中，分析了传染性胃肠炎病毒 (TGEV) 和猪呼吸道冠状病毒 (PRCV) 基因组选定区域的多样性。通过逆转录酶反应和聚合酶链式反应 (RT/PCR) 扩增刺突 (S) 糖蛋白基因的 N 端一半和开放阅读框 (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）和 PRCV 毒株（ISU-1）以及 TGEV 和 PRCV 现场分离株进行了分析。根据 ORF 3、3-1 和 4 RT/PCR 产物的大小，TGEV 和 PRCV 菌株可以快速、轻松地分为三组，即 TGEV Miller、普渡型和 PRCV。通过对S糖蛋白基因N端区域和ORF 3、3-1和4进行RFLP分析，使用限制性酶Sau3AI将TGEV和PRCV菌株区分为五组。对来自强毒力和减毒米勒毒株的 ORF 3、3-1 和 4 中的 PCR 产物进行的序列分析表明，该区域存在其他差异，这些差异与 TGEV 分离株的毒力变化相关。
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.