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采用非放射性标记物－地高辛碱性磷酸酶标记肾综合征出血热病毒（ＨＦＲＳＶ）ｃＤＮＡ制备分子探针，检测ＨＦＲＳＶ在鼠肺、恙螨体内分布定位，并对原位分子杂交的各环节进行探讨。结果表明：鼠肺的吞噬细胞、支气管粘膜上皮细胞、血管内皮细胞等均见有阳性颗粒，在恙螨卵巢细胞、中肠及支囊上皮细胞也存在阳性颗粒；在前处理中，明胶硫酸铬钾与多聚赖氨酸的铺片效果相似；室温２ｈ，４２℃１ｈ，４％多聚甲醛处理冰冻切片，可有效防止脱片，且保持良好的细胞形态结构；探针ＰＣＲ标记法优于随机引物标记法；预杂交液加入ｓｐｅｒｍＤＮＡ和ｙｅａｓｔｔＲＮＡ，不用硫酸萄聚糖，可有效地减少背景染色；４２℃杂交１６ｈ，显色７ｈ可得到理想结果
The non-radioactive marker-digoxin alkaline phosphatase-labeled cDNA of hemorrhagic fever with renal syndrome virus (HFRSV) was used to prepare molecular probes to detect the distribution of HFRSV in mouse lung and chigger mites. To explore. The results showed that there were positive granules in phagocytes, bronchial epithelial cells and vascular endothelial cells in rat lung, and positive particles in the epithelium of chigger ovary, midgut and mesenchymal cells. In the pretreatment, gelatin-sulfuric acid Chromium-potassium and poly-lysine were similar in the effect of plating. The frozen section was treated with 4% paraformaldehyde at 42 ℃ for 1 hour and at room temperature for 2h, which could effectively prevent delamination and maintain good cell morphology. Random primer labeling method; pre-hybridization solution was added spermDNA and yeasttRNA, without sulfuric acid dextran, can effectively reduce the background staining; 42 ℃ hybridization 16h, color 7h can be the ideal result