Larochelle R, Magar R
Laboratoire d'hygiène vétérinaire et alimentaire, Agriculture et Agroalimentaire Canada, Saint-Hyacinthe, Québec.
J Virol Methods. 1997 Jan;63(1-2):227-35. doi: 10.1016/s0166-0934(96)02131-3.
A non-radioactive in situ hybridization (ISH) method developed recently and an immunohistochemical method, the immunogold silver staining (IGSS), were compared for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) in formalin-fixed paraffin-embedded tissues. Serial sections from 98 tissues, representing different organs from PRRSV experimentally infected pigs, and serial sections from 46 lung tissues from field cases were tested by both methods. Results obtained on tissues from experimentally infected pigs and tissues from field cases demonstrated that ISH was more sensitive than immunohistochemistry. More tissues were positive by ISH compared to IGSS and also a greater number of labelled cells and a stronger signal in stained cells were observed in ISH-treated sections. The ISH method described, using a 254 bp digoxigenin-labelled cDNA probe, is a rapid, highly specific and sensitive detection method which can be used for the diagnosis of PRRSV in routinely fixed and processed tissues.
最近开发的一种非放射性原位杂交(ISH)方法与一种免疫组织化学方法——免疫金银染色(IGSS),被用于比较检测福尔马林固定石蜡包埋组织中的猪繁殖与呼吸综合征病毒(PRRSV)。对来自经PRRSV实验感染猪的代表不同器官的98个组织的连续切片,以及来自现场病例的46个肺组织的连续切片,均采用这两种方法进行检测。在实验感染猪的组织和现场病例的组织上获得的结果表明,ISH比免疫组织化学更敏感。与IGSS相比,ISH检测出更多呈阳性的组织,并且在ISH处理的切片中观察到更多的标记细胞以及染色细胞中更强的信号。所描述的ISH方法,使用一个254 bp的地高辛标记cDNA探针,是一种快速、高度特异且灵敏的检测方法,可用于诊断常规固定和处理组织中的PRRSV。