细菌16SrRNA基因检测在新生儿败血症早期诊断中的临床意义  

Role of 16S rRNA gene in rapid diagnosis of neonatal sepsis

作　　者：张芳[1] 宋力[2] 党力亨[2] 孟英韬[2] 张金婷[3] 

机构地区：[1]天津市儿童医院新生儿科,300074 [2]天津市儿童医院儿科研究所,300074 [3]天津市儿童医院检验科,300074

出　　处：《中华围产医学杂志》2008年第3期169-172,共4页Chinese Journal of Perinatal Medicine

摘　　要：目的分析16SrRNA基因检测在早期诊断新生儿败血症中的临床意义。方法收集临床常见的致病菌菌株37株并提取DNA，对106例临床拟诊为败血症的患儿于入院后24h内采集血标本，提取DNA，在16SrRNA基因的保守区选择一对通用引物进行聚合酶链反应（polymerase chain reaction，PCR），并检验实验方法的灵敏性和特异性；106例临床拟诊为败血症的患儿同时查血培养、非特异性炎症指标及降钙素原（procalcitonin，PCT），并采用Χ^2检验与同期住院的20例非败血症患儿进行比较。结果所有细菌PCR均得到预期的约371bp大小的扩增产物，与病毒及人基因组DNA无交叉反应，其扩增下限为10CFU／ml的大肠埃希菌。106例拟诊败血症患儿血培养阳性15例，PCR阳性36例。PCR的敏感性、特异性及诊断指数分别为82．9％、96．9％和179．85，优于血培养及5项非特异指标至少2项异常的实验诊断方法；在围产儿中，PCR的特异性高于PCT。结论PCR方法检测细菌16SrRNA基因能迅速判断临床标本中是否存在细菌，对于早期诊断新生儿败血症具有较高的敏感性及特异性，对于区分败血症与局部感染和非细菌性感染具有重要意义。Objective To investigate the clinical value of 16S rRNA gene in rapid diagnosis of neonatal sepsis. Methods The DNA of 37 strains of common pathogenic bacteria was amplified by polymerase chain reaction （PCR） with a pair of universal primers synthesized based on the highly conservative sequence of bacterial 16S rRNA gene, and the specificity and sensitivity of the PCR were tested. Blood samples from 106 cases of suspected sepsis neonates were collected. The DNA, blood culture, non-specific inflammatory index and procalitonin （PCT） of these neonates were tested and compared with 20 non-sepsis cases with Χ^2-test. Results The 371 bp DNA fragment was amplified successfully from all bacteria strains, which had no cross-reactions with human DNA or virus DNA. Of 106 neonates with suspected sepsis, 15 were positive for blood culture and 36 for PCR. The sensitivity, specificity and diagnosis index of PCR were 82. 9%, 96. 9% and 179. 85 respectively, which were better than those of blood culture and non-specific criteria. Compared with PCT, the specificity of PCR was significantly higher in perinatal neonates. Conclusions 16S rRNA gene amplified by PCR could be used to early detect the pathogenic bacteria in the specimens. It has higher sensitivity and specificity in diagnosis of neonatal sepsis as well as high value in distinguishing it from localized infection and non-bacterial infection.

关 键 词：RNA 核糖体 16S 基因 细菌 败血病 婴儿 新生 早期诊断 

分 类 号：R722[医药卫生—儿科]