通过逆转录聚合酶链反应检测传染性胃肠炎病毒并与猪呼吸道冠状病毒进行鉴别。
Detection of transmissible gastroenteritis virus by RT-PCR and differentiation from porcine respiratory coronavirus.
作者信息
Paton D, Ibata G, Sands J, McGoldrick A
机构信息
Virology Department, Central Veterinary Laboratory (Weybridge), Surrey, UK.
出版信息
J Virol Methods. 1997 Jul;66(2):303-9. doi: 10.1016/s0166-0934(97)00055-4.
Abstract
An RT-PCR method was developed that amplified genetic material from the 5' end of the S protein gene of both transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), but discriminated between the two by the size of the product generated. A number of restriction endonuclease enzymes were assessed for recognition of the amplicons so produced. The assay was shown to detect viral RNA from all of the 26 different TGEV and PRCV isolates examined, covering a period from 1946 to 1996. Detection of TGEV in clinical specimens was possible using a spin column method to extract RNA and sensitivity was compared to virus isolation and antigen detection ELISA. The method could provide a means of confirming positive results from immunological screening tests such as FAT and ELISA, reducing the need for virus isolation and convalescent serology.
摘要
开发了一种逆转录聚合酶链反应（RT-PCR）方法，该方法可扩增传染性胃肠炎病毒（TGEV）和猪呼吸道冠状病毒（PRCV）S蛋白基因5'端的遗传物质，并通过所产生产物的大小区分这两种病毒。评估了多种限制性内切酶对如此产生的扩增子的识别情况。该检测方法可检测1946年至1996年期间检测的所有26种不同的TGEV和PRCV分离株的病毒RNA。使用旋转柱法提取RNA可检测临床标本中的TGEV，并将其灵敏度与病毒分离和抗原检测ELISA进行比较。该方法可为确认免疫筛查试验（如荧光抗体试验（FAT）和ELISA）的阳性结果提供一种手段，减少病毒分离和恢复期血清学检测的需求。
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J Virol Methods. 1996 Feb;56(2):149-60. doi: 10.1016/0166-0934(95)01901-4.
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