胆汁纳米细菌的培养  

Culture of nanobacteria from human gallbladder bile

作　　者：王利民[1] 沈文律[2] 张士莲 曹立赢 王宝强 

机构地区：[1]唐山工人医院普通外科,河北省唐山市063000 [2]汕头大学医学院附属第二医院普通外科,广东省汕头市515041 [3]开滦矿务局医院,河北省唐山市063000

出　　处：《世界华人消化杂志》2005年第22期2663-2666,共4页World Chinese Journal of Digestology

摘　　要：目的:介绍从胆汁中培养纳米细菌的方法. 方法:无菌抽取30例胆囊结石患者的胆汁经“稀释-离心-过滤”或“过滤法”预处理后,在常规细胞培养条件下进行纳米细菌培养．光镜下观察细菌形态．采用间接免疫荧光染色对培养所得细菌进行鉴定,并在电镜下观察．结果:30例胆汁样本,经过“稀释-离心-过滤”预处理后,纳米细菌培养的阳性率为40％,经“过滤法”预处理后,纳米细菌培养的阳性率为57％,两种方法预处理后,胆汁纳米细菌培养阳性率无统计学差异(x2=1．669, P>0．05)．培养2 wk时,显微镜下可见做布朗运动的微小的颗粒状纳米细菌．4 wk时,可见纳米细菌开始贴附于培养瓶底部形成细菌被膜．培养所得纳米细菌与8D10 抗体产生特异性结合,电镜下观察呈球形或短棒状颗粒,大小约为80-350 nm．结论:在胆囊结石患者的胆汁中存在纳米细菌感染, “过滤法”是一种简单有效的预处理胆汁的方法．AIM： To introduce a method for culturing nanobacteria from human gallbladder bile. METHODS： The bile samples were collected from 30 patients with gallbladder-stone. After pretreatment with dilution-centrifuge-filtration （DCF） and filtration method, respectively, those samples were cultured under the routine cell culture condition for nanobacteria. The morphological features of the obtained nanobacteria were observed under light and electron microscope. Indirect immunofluorescence staining was used to identify the nanobacteria. RESULTS： Of the 30 bile samples, the positive rate of nanobacteria was 40% when the samples were pretreated by DCF method, and the positive rate was 57% when they were pretreated by filtration method. There was no significant difference between those two methods （χ^2 = 1.669, P 〉0.05）. Brown movement of the tiny nanobacteria was observed at 2 wk. At 4 wk, nanobacteria started to attach on the bottom of the culture bottles. Specific combination of the obtained nanobacteria with 8D10 antibodies was observed. The nanobacteria were ball- or stick-like with a length of 80-350 nm under electron microscope. CONCLUSION： The infection of nanobacteria exists in the bile of gallbladder-stone patients. Filtration is a simple and effective method for the preparation of the gallbladder bile.

关 键 词：胆囊结石 纳米细菌 细胞培养 电子显微镜 间接免疫荧光染色 

分 类 号：R575.6[医药卫生—消化系统]