胆汁纳米细菌的培养与鉴定

目的：探讨纳米细菌在胆汁中的分布情况及微生物学特征,并对其鉴定方法进行评价。方法：对75例胆囊胆汁标本进行纳米细菌培养,观察其生长情况,并采用免疫组化染色、透射电镜扫描和钙染色进行纳米细菌鉴定。结果：胆汁中纳米细菌感染率为61．3％,其中出现完全或部分黏附于管底壁的白色沉淀者,纳米细菌阳性率为75．7％,而沉淀完全呈絮状或无沉淀者,阳性率为47．4％,两者间存在显著性差异。免疫组化染色阳性的胆汁标本经透射电镜扫描均发现纳米细菌。钙染色的阳性率为38．7％,与免疫组化染色相比较,其灵敏度为58．7％,特异度为93．1％,假阳性率6．9％,假阴性率41．3％。结论：纳米细菌单克隆抗体联合透射电镜扫描可鉴定纳米细菌;钙染色对纳米细菌的鉴定有着重要的参考价值;紧密黏附于管底的白色沉淀是纳米细菌培养过程中的一个重要微生物学特性。

Culture and identification of nanobacteria in bile

OBJECTIVE: To study the distribution and identification of nanobacteria in bile and to evaluate the identifying methods of nanobacteria. METHODS: RPMI1640 culture or RPMI1640 culture with 10% heat-inactivated gamma-FBS was added into 75 samples of cystic bile from gallbladders resected in operation. Nanobacteria were identified by immunohistochemical staining, transmission electron microscopy (TEM), and calcific staining. RESULTS: Nanobacteria were found in 45 bile samples with a positive rate of 61.3%. The positive rate of nanobacteria was 75.7% among 37 bile samples with white precipitate adhering to the tube, and was 47.4% among the samples with flocculent precipitate or without precipitate (P < 0.05). The immunohistochemically confirmed presence of nanobacteria was re-confirmed by TEM in all the positive samples. The positive rate, sensitivity, specificity, false positive rate and false negative rate of calcific staining were 38.7%, 58.7%, 93.1%, 6.9% and 41.3% respectively. CONCLUSION: Immunohistochemistry with monoclonal antibody of nanobacteria associated with TEM is useful in identifying nanobacteria. Calcific staining is of great value to identification of nanobacteria. Precipitation of white floccules adhering to the tube is an important microbiological characteristic of nanobacteria.