基因检测在新生儿脓毒症早期诊断中的价值  

Clinical Value of Gene Detection in Early Diagnosis of Sepsis in Neonates

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

机构地区：[1]天津市儿童医院新生儿科,天津300074

出　　处：《实用儿科临床杂志》2011年第10期764-766,共3页Journal of Applied Clinical Pediatrics

摘　　要：目的分析16SrRNA基因检测在新生儿脓毒症早期诊断中的临床价值，探讨快速、可靠地诊断该症的试验方法。方法在16SrRNA基刚果守区选择一对通用引物，收集临床常见的致病菌株37株，提取细菌DNA并进行PCR检测，采用琼脂糖凝胶电泳法观察扩增结果，同时对该引物的灵敏性及特异性进行检验。对106例临床疑诊为脓毒症的新生儿于入院24h内，在严格无菌条件下采集血标本，提取DNA进行PCR扩增，同时行血培养、血常规检查及CRP及ESR水平检测，并与同期住院的20例非感染性疾病患儿进行比较。结果PCR检测细菌DNA均得到预期的约371bp大小的扩增产物，该引物仅扩增细菌DNA，与病毒及人基因组DNA无交叉反应，其扩增下限为10^4CFU·L^-1的大肠埃希菌。106例疑诊为脓毒症的患儿，血培养阳性15例，PCR检测阳性36例，PCR的阳性检出率显著高于血培养（P〈0．005），20例非感染组患儿PCR结果均为阴性。依据新生儿脓毒症诊断标准，PCR的敏感性、特异性及诊断指数分别为82．93％、96．92％和179．85，优于血培养及5项非特异指标至少2项异常的诊断方法。PCR方法6—8h即能检测出结果，改良核酸提取技术可以将检测时间缩短至4～6h。结论PCR方法检测细菌16SrDNA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脓毒症具有较高的敏感性及特异性。Objective To analyze the clinical value of 16S rRNA gene in early diagnosis of neonatal sepsis and find out a rapid and accurate assay to diagnose it.Methods A universal primer was synthesized based on the highly conserved sequences of bacterial 16S rRNA gene,37 strains of common pathogenic bacterium were amplified by polymerase chain reaction（PCR） and the result was observed with agorose gel electrophoresis.The specifity and sensitivity of the PCR were tested.Next,blood speciments from 106 cases of suspected sepsis were cultured and DNA isolated from these speciments was amplified by PCR also,C reactive protein,blood routine and erythrocyte sedimentation rate were detected,and compaired with 20 non-infected cases.Results The positive fragment with length of 371 bp were amplified from all strains.The primer could only amplify bacteria no cross reaction with human DNA and viruses DNA,and its sensitivity could be improved to 104CFU·L-1.Of 106 speciments with suspected bacterial,15 cases were positive on blood culture and 36 cases on PCR.The positive rate of PCR was significantly higher than that of blood culture（P0.005）,while 20 specimens of non-infected infants were all negative on PCR.The sensitivity,specifity and diagnosis index of PCR were 82.93%,96.92% and 179.85,respectively,which were superior to blood culture and non-specific criteria.The time of PCR spent only for 6-8 h,improving the extracting technique of DNA,it could be 4-6 h.Conclusions 16S rDNA amplified by PCR can be used to detect the pathogenic bacteria in specimens rapidly.It has higher sensitivity and specificity in diagnosis of neonatal sepsis.

关 键 词：16S 核糖体核糖核酸 细菌 脓毒症 婴儿 新生 

分 类 号：R722.1[医药卫生—儿科]