肾综合征出血热患者尿融合细胞内病毒G区基因及其蛋白表达

Detection of G gene and its expression of hantaviruses in the urine fusion cells of hemorrhagic fever with renal syndrome

目的：研究肾综合征出血热（ＨＦＲＳ）患者尿融合细胞内汉坦病毒（ＨＶ）Ｇ区靶基因（膜蛋白基因）ＲＮＡ、ｍＲＮＡ的定位和ＨＶ膜蛋白在尿融合细胞中的表达定位。方法：设计针对病毒Ｍ片段Ｇ基因区的探针，地高辛素标记，运用核酸原位杂交技术检测靶基因ＲＮＡ 和ｍＲＮＡ在尿融合细胞中的定位。同时用ＳＡＢＣ免疫组化法检测病毒膜蛋白在尿融合细胞中的表达定位。结果：病毒靶ＲＮＡ和ｍＲＮＡ检出于细胞核和细胞浆。２１例中，１４例阳性，阳性率６６．６％（１４／２１），其中核型９例，核浆混合型５例。病毒膜蛋白检测，２１例中１６例阳性，阳性率７６．２％（１６／２１），病毒膜蛋白表达见于细胞膜。两者阳性率相比较差异无显著性（Ｐ＞０．０５）。结论： ＨＦＲＳ病毒膜蛋白的表达与病毒编码该蛋白的Ｇ基因区ＲＮＡ、ｍＲＮＡ均可在患者尿融合细胞内检出。

Objective:To localize G gene and its expression o f Hantaviruses in the urine fusion cells of patient with HFRS.Methods:The probes were synthesized,the sequences of which were based on those of Hantavirus nucleic acid M fragment G gene, and labeled with Digoxigenin on terminal 5'. They were used to loc alize the target gene RNA or mRNA in the urine fusion cells, by in situ hybridiz ation. Streptavidin biotin complex immunohistochemical assay was used to detect the glycoprotein expressed on the urine fusion cells. Results:Hantavirus nucleic acid M fragment G gene RNA or mRN A were localized in the nuclei or cytoplasm of the fusion cells. The positive rate of in situ hybrid ization detection was 66.6%(14/21). Among 14 positive cases, 9 were karyon t ype and 5 were mixed type of karyon and cytoplasm. Hantaviruses glycoprotein was de te cted on or in the urine fusion cells, mainly expressed on the membrane. The pos itive rate of immunohistochemical detection was 76.2%(16/21). There was no difference between the two positive rates (P>0.05) .Conclus ion:Simple and accurate in situ hybridization assay can be used reliably to localize the target gene in the urine fusion cells；Hantavirus es glycoprotein expressed on the fusion cells corresponds to the gene RNA or mRNA of M fragment G gene in the fusion cells.