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既往认为HBV具有严格的嗜肝性。但最近的研究表明,HBV感染不仅限定于肝脏,而且可在人体多种组织器官中存在。我们采用分子杂交技术,对3例乙肝患者尸检部分器官进行了观察。 材料和方法 1.尸检对象,为3例乙肝患者和l例非乙肝患者(做为对照),详见表1。 2.方法:(1)标本处理:取适量冻存组织块,用 PBS洗去表面物,剪成2-3mm小块。磷酸盐缓冲盐水溶液(PBS)洗3次,经胰蛋白酶消化后,洗去胰酶。加少量缓冲液,用吸管吹打使细胞分散。再用0.14MNaCl溶液洗3次,收集细胞沉淀物。(2)DNA提取;参照顾健人的方法(中华传染病杂志1984,4(2):221)。(3)斑点杂交:参照杜绍财的方法[上海免疫学杂志1084;4(4):230]。 结果:显示黑色斑点为阳性,详见表2。 表2 尸检组织器官HBV—DNA检测结果
In the past that HBV has a strict hepatic. However, recent studies have shown that HBV infection is not limited to the liver, but also in many tissues and organs of the human body. We used molecular hybridization technique to observe the autopsy of some organs in 3 hepatitis B patients. Materials and methods 1. The subjects were autopsy, 3 cases of hepatitis B patients and l cases of non-hepatitis B patients (as a control), as shown in Table 1. 2. Methods: (1) specimen processing: take the appropriate amount of frozen tissue block, wash the surface with PBS, cut into 2-3mm small pieces. Phosphate buffered saline (PBS) wash 3 times, trypsinized, wash away trypsin. Add a small amount of buffer and pipette to disperse the cells. Wash with 0.14M NaCl again three times and collect cell pellet. (2) DNA extraction; refer to Gu Jianren’s method (Chinese Journal of Infectious Diseases 1984,4 (2): 221). (3) Spots hybridization: refer to the method of Du Shaocai [Shanghai Immunology 1084; 4 (4): 230]. Results: Black spots are positive as shown in Table 2. Table 2 autopsy tissues and organs HBV-DNA test results