【摘要】 目的探讨能否使用咽拭子培养结果推测细菌感染性肺炎的病原菌。方法选取2013年11月至2014年2月因患严重细菌感染性肺炎在重庆医科大学附属儿童医院新生儿重症监护室住院并需机械通气辅助治疗的37例新生儿作为研究对象。在插管3 d内至少采集一次痰液并分为2份,一份做培养,另一份存-20℃冰箱备检。在收集痰液的同时采集咽拭子。对痰液培养出致病菌患儿的另一份痰液和对应咽拭子进行如下操作:从痰液中提取细菌DNA,经16S r DNA PCR扩增和变性梯度凝胶电泳(DGGE)后将分离出的DNA进行克隆和测序,通过美国国立生物技术信息中心(NCBI)数据库进行Blast比对,同时分析菌群的组成。结果上、下呼吸道细菌种类和每个菌属的构成比比较,差异均有统计学意义(P<0.05),其中7个菌属是相同的。同时,培养出的肺炎克雷伯菌、鲍曼不动杆菌和铜绿假单胞菌所属的克雷伯菌属、不动杆菌属和假单胞菌属正好是上、下呼吸道所共有的菌属。结论如果对咽拭子进行培养,且培养出的致病菌是上、下呼吸道所共有,则提示该菌可能是细菌感染性肺炎的致病菌。更多还原

【Abstract】 Objective To explored whether the culture results from the pharyngeal swabs could be used to determine the pathogens of the neonatal pneumonia. Methods Thirty-seven neonates suffered from severe bacterial pneumonia and require mechanical ventilation in Neonatal Intensive Care Unit of Children′ s Hospital of Chongqing Medical University from November2013 to February 2014 were taken as the research subjects.Within three days after intubation,the sputum was collected at least once and divided into two portions,one for culturing and the other stored at-20 ℃ as a backup sample. Meanwhile,a pharyngeal swab was collected. For neonates with pathogens in the cultures of their sputa,the following procedures were conducted using the backup sputum sample and the corresponding pharyngeal swab. The DNA extracted from the samples was amplified by 16 S r DNA PCR and separated by denaturing gradient gel electrophoresis(DGGE),then performed with gene cloning and sequencing. The resulting sequences were compared with the data in the National Center for Biotechnology Information(NCBI) Gen Bank by using Blast and the composition of the flora was analyzed. Results The difference of bacterial genera and composition ratio of each bacterial genus from the upper and lower respiratory tracts were stastically significant(P<0.05). Seven common genera in both the upper and lower respiratory tracts. The cultured bacteria Klebsiella pneumoniae subspecies,Acinetobacter baumannii and Pseudomonas aeruginosa belonged to the Klebsiella sp.,Acinetobacter sp. and Pseudomonas sp.,respectively,which were the common genera of the upper and lower respiratory tracts. Conclusion It suggests that if the cultured pathogenic bacteria is common to both upper and lower respiratory tracts after culturing of pharyngeal swabs,the cultured bacteria might be the pathogenic bacteria to neonatal bacterial pneumonia.更多还原