用原位RT-PCR分子杂交定位检测螨(革螨、恙螨)原代培养细胞内HV-RNA的研究(Ⅱ)

目的　研究革螨、恙螨在传播肾综合征出血热病毒 (Hemorrhagicfeverwithrenalsyndromevirus,HFRSV)中的媒介作用。方法　采用革螨、恙螨饲养繁殖的子代幼虫、若虫和成虫 ,消化后作螨细胞原代培养制螨细胞片 ,用原位RT -PCR分子杂交法检测HV -RNA在螨组织细胞内分布和定位。结果　HV -RNA多分布于螨消化系统的上皮细胞 ,卵巢组织细胞内 ;革螨子 4代、子 3代和恙螨若虫组织细胞内HV阳性颗粒较革螨子 2代、子 1代和恙螨幼虫多且密集。结论　革螨、恙螨作为HFRS的生物学媒介有细胞分子水平的直接证据。

Detection and location of HV-RNA in primary cultured mite cells by in situ PCR

Aim To study the vector effect of gamasid mites and chigger mites in transmitting the virus of hemorrhagic fever with renal syndrome (HFRSV) Methods The cells of gamasid mites and chigger mites at larva,nymph and adult stages were cultured,HV RNA in primary cultured cells were examinated by situ RT PCR Results The results showed that the positive signals of HV RNA distributed mostly in epithelial cells of digestive system and ovary cells of larva and nymph The signal particles in the third and fourth generation cells of gamasid mites and nymph cells of chigger mites were more than that in the first and second generation cells of gamasid mites and larva cells of chigger mites Conclusion The gamasid mite and chigger mite as the biological vector of HFRS have the direct evidence on the cell and molecular level