16S rRNA基因及多重聚合酶链式反应在新生儿败血症早期诊断中的应用研究  

Use of multiplex-PCR targeting 16S rRNA gene for early diagnosis of neonatal sepsis

作　　者：秦桂秀[1] 黄瑞[1] 张莉[1] 孟晋华[1] 李文玲[1] 郭慧敏[1] 朱镭[1] 

机构地区：[1]山西省儿童医院检验科,太原030013

出　　处：《中国药物与临床》2017年第11期1566-1568,共3页Chinese Remedies & Clinics

基　　金：山西省基础研究计划项目(2013011059-5)

摘　　要：目的分析16S rRNA基因聚合酶链式反应(PCR)检测方法及血培养方法检测疑似新生儿败血症患儿血标本中潜在病原菌的病原学特点,并以此为基础建立一套适用于临床的可检测新生儿败血症常见菌株的特异性引物多重PCR体系。方法建立16S rRNA通用引物PCR检测方法并检测534例疑似新生儿败血症患儿血标本,分析其病原学感染及特点,与常规血培养检测结果进行比较。建立适用于临床的可检测新生儿败血症常见菌株的特异性引物多重PCR体系。结果 534例血标本的血培养阳性率为26%,经鉴定为凝固酶阴性葡萄球菌(44.6%)、大肠埃希菌(14.4%)、无乳链球菌(8.6%)、金黄色葡萄球菌(7.9%)、肺炎克雷伯菌(5.8%)、屎肠球菌(4.3%)、其他菌株(14.4%)。16S rRNA通用引物PCR检测阳性率为37.6%,显著高于血培养检测(P<0.05)。经优化的多重PCR体系可快速准确检测败血症常见菌种。结论 16S rRNA基因PCR方法用于新生儿败血症的检测具有较高的检出率,可用于临床新生儿败血症的快速诊断。本研究初步建立的败血症常见致病菌特异性引物多重PCR体系具有一定的临床应用价值。Objective To analyze the characteristics of potential pathogens in blood samples from children with suspected neonatal sepsis by using multiplex-PCR targeting 16 S rRNA gene compared with blood culture, and accordingly, set up a multiplex PCR system with specific primers which is clinically useful for detection of common bacterial strains in neonatal sepsis. Methods PCR with 16 S rRNA universal primers were used to detect the blood samples from 534 children with suspected neonatal sepsis. Findings of the pathogens and characteristics of the infection were analyzed and compared with those from conventional blood culture. A multiplex PCR system with specific primers was subsequently developed for clinical detection of common bacterial strains in neonatal sepsis. Results By blood culture, the positive rate was 26% for pathogens in the 534 blood samples, which were identified to be coagulase-negative staphylococci（44.6%）, Escherichia coli（14.4%）, Streptococcus agalactiae（8.6%）, Staphylococcus aureus（7.9%）,Klebsiella pneumoniae（5.8%）, Enterococcus faecium（4.3%）, and other strains（14.4%）. The positive rate of PCR with16 S rRNA universal primers was 37.6%, which was significantly higher than that of blood culture（ P〈0.05）. The optimized multiplex PCR system can detect the common bacteria in sepsis quickly and accurately. Conclusion PCR targeting the 16 S rRNA gene may yield a high detection rate for pathogens in neonatal sepsis and can be used clinically for the rapid diagnosis of this condition. The multiplex PCR system with pathogen-specific primers preliminarily established in this study for detecting common pathogens in sepsis may have a role in clinical practice.

关 键 词：基因 rRNA 培养技术 多重聚合酶链式反应 

分 类 号：R440[医药卫生—诊断学] R722.131[医药卫生—临床医学]