胆结石与胆汁中纳米细菌的检测与意义  

Significance of detection of nanobacteria in bile and gallstones

作　　者：朱明利 王晶晶 潘万能 刘寿荣 朱元东 ZHU Ming-li;WANG Jing-jing;PAN Wan-neng;LIU Shou-rong;ZHU Yuan-dong(Hangzhou Xixi Hospital,Hangzhou,Zhejiang 310023,China;不详)

机构地区：[1]杭州市西溪医院,浙江中医药大学附属杭州西溪医院开放实验室,浙江杭州310023 [2]浙江萧山医院感染病科,浙江萧山311202

出　　处：《中华医院感染学杂志》2019年第21期3279-3283,共5页Chinese Journal of Nosocomiology

基　　金：杭州市卫生科技计划重点基金资助项目(2010Z008)

摘　　要：目的研究胆结石患者胆汁及结石中纳米细菌(NB)的感染情况,分析纳米细菌与胆结石形成的关系,探讨纳米细菌在胆结石形成中的可能机制。方法选择2013年10月-2014年12月杭州市西溪医院外科手术患者,术中无菌收集胆汁43例(胆囊结石患者36例,非胆囊结石患者7例),结石30例。采用细胞培养的方法,对胆汁及结石中的纳米细菌进行分离培养。采用间接免疫荧光法、透射电镜负染法、茜素红钙染色法、PCR法等对纳米细菌进行鉴定。结果以间接免疫荧光法为金标准,胆汁中纳米细菌阳性率为44.18%(19/43),其中胆结石患者胆汁纳米细菌阳性率为50.00%(18/36),非胆囊结石患者胆汁中纳米细菌阳性率为14.29%(1/7);结石中纳米细菌阳性率为60.00%。胆囊结石患者胆汁与结石中NB的阳性率无统计学差异。钙染色法与间接免疫荧光法、PCR法与间接免疫荧光法检测纳米细菌无统计学差异。结论纳米细菌与胆结石密切相关。胆结石患者纳米细菌的感染率较高,可能会促进胆结石的形成。采用细胞培养、稀释双过滤的方法能从胆汁及结石中分离培养出纳米细菌;透射电镜负染法、间接免疫荧光法可作为纳米细菌鉴定的金标准,茜素红钙染色、PCR法可作为重要参考。OBJECTIVE To observe the prevalence of nanobacteria(NB)infection in bile and calculi of the patients with gallstones,analyze the correlation between the nanobacteria and formation of gallstone and explore the mechanisms of formation of gallstones so as to provide guidance for prevention and treatment of gallstones.METHODS The patients who received surgeries in Hangzhou Xixi Hospital from Oct.2013 to Dec.2014 were enrolled in the study.The bile specimens were collected from 43 patients including 36 gallstone patients and 7 non-gallstone patients and 30 patients with calculi during surgery.The bile and calculi specimens were cultured for NB by using cell culture method,and the NB strains were identified with the use of indirect immunofluorescence staining,transmission electron microscopy,alizarin red calcium staining and PCR.RESULTS Set the indirect immunofluorescence staining as the gold standard,the positive rate of NB was 44.18%(19/43)in the bile specimens,the positive rate of NB was 50.00%(18/36)in the bile specimens of the gallstone patients,14.29%(1/7)in the bile specimens of the non-gallstone patients.The positive rate of NB was 60.00%in the calculi specimens;there was no significant difference in the positive rate of NB between the bile and calculi specimens of the gallstone patients.There was no significant difference in the detection of NB between the calcium staining method and the indirect immunofluorescence staining method and between the PCR and the indirect immunofluorescence staining method.CONCLUSION The NB is closely associated with gallstones.The incidence rate of NB infection is high among the gallstone patients,which may accelerate the formation of gallstones.The NB strains can be isolated from the bile and calculi specimens by cell culture and dilution double filtration method.Transmission electron microscopy and indirect immunofluorescence staining can be used as gold standards for identification of NB,and alizarin red calcium staining and PCR can provide significant reference.

关 键 词：胆囊结石 胆汁 纳米细菌 透射电镜负染法 间接免疫荧光法 

分 类 号：R575.6[医药卫生—消化系统]