摘要
目的探讨本地区儿童肺炎细菌性病原分布,指导肺炎病原临床诊治及合理用药。方法收集确诊为社区获得性肺炎患儿呼吸道分泌物188例,细菌培养并提取其DNA,多重PCR同时扩增14种呼吸道病原菌靶基因,扩增产物采用Luminex 100点阵仪检测,比较多重PCR检测敏感性和特异性;分析肺炎细菌病原混合感染情况。结果 188例呼吸道标本中,细菌培养出114株病原菌,阳性率为60.6%(114/188)。经多重PCR扩增、Luminex 100点阵仪检测75例呈阳性,阳性率为39.9%(75/188)。多重PCR对14种病原菌敏感性为51.0%,特异性为68.0%,符合率为58.3%。多重PCR检测肺炎链球菌检出率(19.1%,36/188)高于细菌培养(3.7%,7/188)(P<0.01);联合检测细菌性病原检出率为78.2%(147/188),联合检测提高流感嗜血杆菌、肺炎链球菌、金黄色葡萄球菌、铜绿假单胞菌和鲍曼不动杆菌检出例数。肺炎支原体与细菌混合感染16例,混合感染率为8.5%(16/188),最常见是肺炎支原体与流感嗜血杆菌混合感染,占混合感染的50.0%(8/16)。结论多重PCR技术对肺炎链球菌有很高的敏感性,细菌培养和病原特异性DNA联合检测能提高儿童肺炎细菌性病原检出率,应用多重PCR可以及时明确CAP病原中肺炎支原体与细菌混合感染。
Objective To approach of multiplex PCR for the pneumonic bacterial etiologic diagnosis in littoral of Zhejiang,direct etiologic diagnosis and antibiotic rational use,and understand the distribution of bacterial etiology. Methods 188 hospitalized children in our hospital,were collected deep tracheal aspirate at the time of hospitalization. The respiratory tract secretions were immediately sent for bacterial culture. Then we extracted DNA from secretions. Simultaneously,14 respiratory tract pathogenic bacterium were detected by multiplex PCR. Amplification products were identified by the Luminex 100 suspension array. The sensitivity and specificity of multiplex PCR for the bacteria were evaluated. The affect for pneumonic bacterial etiologic distribution was analysised by the combined detection with bacterial culture and multiplex PCR assay. Results Of the 188 respiratory tract specimens, 114 were positive by bacteria culture. The positive rate of culture was 60. 6%( 114 /188). The target gene were detected by multiplex PCR from 75 specimen,the positive rate of multiplex PCR was 39. 9%( 75 /188). For the 14 kinds of bacterium designed by multiplex PCR,compared with the culture,the sensitivity,specificity and coincidence of multiplex PCR weres 51. 0%,68. 0% and 58. 3%,respectively. Multiplex PCR was more sensitive than culture in the detection of Streptococcus penumoniae( 36 /188 19. 1% vs 7 /188 3. 7%,P < 0. 01). The total positive rate by combined detection of culture and multiplex PCR assay was 78. 2%( 147 /188). The combined detection may increase the positive case of Staphylococcus aureus, Hemophilus influenzae, Staphylococcus aureus, Pseudomonas aeruginosa and Acinetobacter baumannii. The mixed infection with Mycoplasma pneumoniae and bacterium was 16 by multiplex PCR. Of which,the most common mixed infection was Mycoplasma pneumoniae and Hemophilis influenzae,it was 50. 0%( 8 /16). Conclusion Multiplex PCR is highly sensitive for detection of Streptococcus penumoniae causing pneumonia in children. The combined detection of bacterial culture and multiplex PCR assay may increase the positive rate of bacterial pathogens in hospitalized children with CAP and can really demonstrate the distribution of bacterial etiology. Multiplex PCR can timely indicate the mixed infection with Mycoplasma pneumoniae and bacterium in etiologic agent causing CAP.
出处
《中国微生态学杂志》
CAS
CSCD
2014年第1期46-50,共5页
Chinese Journal of Microecology
基金
温州市科技计划项目(Y20130076)
关键词
聚合酶链反应
肺炎
病原学
Polymerase chain reaction
Pneumonia
Etiology
