慢性肝病和肝癌患者血液中纳米细菌的检测  

Detection of nanobacteria in blood of patients with chronic hepatopathy and hepatocellular carcinoma

作　　者：朱明利[1] 娄国强[1] 罗英 李超丹[1] 厉小玉[1] 刘寿荣[1] 周俊[1] 胡林 

机构地区：[1]杭州市第六人民医院浙江中医药大学附属杭州第六医院开放实验室,杭州310014 [2]杭州优思达生物技术有限公司

出　　处：《中国人兽共患病学报》2008年第7期621-625,630,共6页Chinese Journal of Zoonoses

基　　金：杭州市医药卫生科技计划重点项目(2003Z007)

摘　　要：目的研究纳米细菌的分离与鉴定方法,了解慢性肝病和肝癌患者血液纳米细菌(NB)感染状况。方法对68例慢性乙型肝炎(CHB)、54例慢性重型乙型肝炎(CSHB)、66例肝硬化(CL)、23例肝癌(HCC)患者采用培养、免疫组化、钙染色,检测血液中纳米细菌,并与40例健康人结果比较。部分培养物用透射电镜观察。结果CHB、CSHB、CL、HCC病人和健康人血清纳米细菌免疫组化阳性率分别为23.91%、36.84%、38.89%、26.09%和7.50%。钙染色分别为29.55%、45.45%、37.14%、18.18%和5.00%,肝病阳性率均高于健康人(P<0.05)。显微镜下,纳米细菌阳性组出现针状、巢装集落或结晶者为82.35%,而阴性者40.00%,两者间差异有统计学意义(P<0.05)。免疫组化阳性的培养物透射电镜可见纳米细菌样微生物。结论单克隆抗体免疫组化联合透射电镜、钙染色可用于纳米细菌的检测。部分慢性肝病、肝癌病人存在血液纳米细菌感染,肝病患者血液纳米细菌感染率高于健康人。To investigate the methods of isolation and identification for nanobacteria（NB） and to explore the status of infection with nanobacteria in blood of patients with chronic hepatopathy and hepatocellular carcinoma, the nanobacteria in blood of 68 patients with chronic viral hepatitis B（CHB）, 54 patients with severe chronic hepatitis B（CSHB）, 66 patients with cirrhosis of liver（CL）and 23 patients with hepatocellular carcinoma（HCC）were investigated by methods of bacteria cultivation, immunohistochemical staining（IHC）and calcium stalning,and the results were compared with those of 40 healthy adults. Some positive culture samples were re-confirmed with the transmission electron microscopy（TEM）. As demonstrated by immunohistochemical stalning,the infection rates with nanobacteria for healthy adults,CHB,CSHB,CL and HCC patients were 7. 5%, 23.91%, 36.84 %, 38.89 % and 26.09 % respectively, whereas, as demonstrated by calcium staining, these figures would be 5 % for healthy adults,29.55% for CHB,45.45% for CSHB,37.14% for CL and 18.18% for HCC. All-together, the positive rates of infections with nanobacteria were significantly higher than those of the healthy adults（P〈0 05）. For NB-IHC-positive group, the positive rates of cultures with needle-like clusters or apatite crystals as observed under microscope was 82.35 % ,but it was 40 % in NB-IHC-negative group ; significant difference existsed between these two groups（P〈0.05）. In addition, ellipse or round shaped microorganisms with diameter of 50-500 nm were found under TEM. These results indicate that immunohistochemical staining together with transmission electronmicroscopy and calcium staining can be used for the detection of nanobacteria and the occurrence of nanobacteria in blood of patients with hepatopathy and hepatocellular carcinoma is higher than that of the healthy adults.

关 键 词：纳米细菌 慢性乙型肝炎 肝硬化 肝癌 

分 类 号：R446.5[医药卫生—诊断学]