摘要
背景泪囊炎是眼科常见的感染性疾病，对泪囊炎致病菌检测的金标准为培养法，在培养法的基础上结合分子生物学的检测方法可以更精确地鉴定出致病菌的种属，但目前眼科领域类似的研究资料比较缺乏。目的采用PCR法扩增细菌核糖体16SrRNA基因序列鉴定泪囊炎病原细菌的种属。方法取10例质控标准菌样本，利用PCR的加热过程使细菌核酸释放，扩增出细菌16SrRNA基因序列，测序后与GenBank中的基因序列进行比对，鉴定出细菌的种属信息，与生化鉴定法结果进行对比，确定16SrRNA基因检测方法的可靠性。取30例泪囊炎患者泪囊分泌物培养出的细菌，利用上述方法进行病原菌鉴定。结果10例质控标准菌样本经PCR扩增后其产物序列的鉴定结果与生化鉴定法结果完全一致。30例泪囊炎病原细菌标本中，16SrRNA基因序列法鉴定结果为表皮葡萄球菌13例，沃氏葡萄球菌2例，人葡萄球菌1例，麦氏棒杆菌5例，肺炎链球菌3例，蜡状芽孢杆菌2例，藤黄微球菌1例；卡他莫拉菌1例，奥斯陆莫拉菌1例，铜绿假单胞菌1例。结论通过PCR扩增细菌核糖体16SrRNA基因序列鉴定泪囊炎病原细菌种属的方法准确性高，特异性强。
Background Dacryocystitis is one of the most common infectious eye diseases. The gold standard for the identification of bacteria causing dacryocystitis is bacterial culture. The combination of regular culture method with molecular biology techeniques will generate more reliable results. However, very few research data are available in ophthalmological studies in this area. Objective This study was to identify the genera and species of the dacryocystitis-causing bacteria by PCR amplification of the 16S rRNA sequences. Methods Ten cases of qualified standardized bacteria samples were taken, and the nucleic acids were released in the heating process of the PCR procedure. The 16S rRNA genes were amplified and sequenced,and the genera and species were identified using BLAST from GenBank, and the results were used to compare with the results from biochemical identification to test the reliability of this method. The cultured bacterial species from the lacrimal sac secretions from 30 cases of dacryocystitis patients were identified with the above method. Results The outcome of the PCR identification for the 10 cases of quality control standard bacterial specimens was consistent with the results from the biochemical identification. The identification of the 30 cases of dacryocystitis through sequencing the 16S rRNA revealed there were 13 cases of Staphylococcus epidermidis infection, 2 cases of Staphylococcus warneri infection, 1 case of Staphylococcus hominis infection,5 cases of Corynebacterium macginleyi infection,3 cases of Streptococcus pneumonia infection,2 cases of Bacillus cereus infection, 1 case of Micrococcus luteus infection, 1 case of Moraxella catarrhalis infection, 1 case of Moraxella osloensis infection and 1 case of Pseudomonas aeruginosa infection. Conclusions Sequencing the 16S rRNA is an accurate and specific way for the identification of the genera and species of bacteria that cause dacryocystitis in patients. This sequencing method is feasible in monitoring a variety of dacryocystitis-causing pathogens. More information and epidemiological statistics about dacryocystitis can be obtained from 16S rRNA sequencing.
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2013年第9期867-869,共3页
Chinese Journal Of Experimental Ophthalmology
