RNA原位杂交法检测组织内弓形虫的实验研究

Experimental study on the detection of Toxoplasma on the paraffin section of liver by RNA in situ hybridization

目的 探讨RNA原位杂交法在弓形虫病理检测中的应用。方法 设计并合成弓形虫SAG2基因来源的寡核苷酸探针。弓形虫RH株感染昆明株小鼠 ,分别于感染后第 2、3、4、5、6、7天处死 ,取肝组织制备石蜡标本进行RNA原位杂交 ,结果同HE染色及免疫组化相比较。结果 RNA原位杂交法于感染后第 2天检到阳性结果 ,组织内弓形虫的检出率 (2 8/ 2 8)高于免疫组化 (2 1/ 2 8)及HE染色法 (16 / 2 8)。结论 RNA原位杂交法是一种简便准确的弓形虫病理检测方法。

To investigate the possible application of RNA in situ hybridization for the detection of toxoplasma in the paraffin section of liver in the experimentally infected mice,mice inoculated with the RH strain of Toxoplasma gondii were sacrificed 2,3,4,5,6 and 7 days after infection,and the paraffin sections of liver were prepared to perform the in situ hybridization with Dig-labeled oligonucleotide probe complememtary to SAG2 mRNA of toxoplasma.The results were parallelly compared with those obtained by HE and immunohistochemical staining.It was found that the specific positive results were observed on sections 2 days after infection.All the 28 paraffin sections detected by in situ hybridization demonstrated positive results,while only 21 and 26 sections showed positive results by immunohistochemical and HE staining respectively.It concludes that the RNA in situ hybridization is a simple and precise method for the detection of toxoplasma in pathological sections.