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目的分析16S rRNA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16S rRNA基因保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;;DOI:10.3969/j.issn.NA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24 h内,在严格无菌条件下采集血标本,提取;;DOI:10.3969/j.issn.NA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;;DOI:10.3969/j.issn.NA均得到预期的约371 bp大小的扩增产物,该引物仅扩增细菌;;DOI:10.3969/j.issn.NA,与病毒及人基因组;;DOI:10.3969/j.issn.NA无交叉反应,其扩增下限为104CFU.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 6~8 h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4~6 h。结论 PCR方法检测细菌16S r;;DOI:10.3969/j.issn.NA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 detection in the early diagnosis of neonatal sepsis and to explore a rapid and reliable method for the diagnosis of the disease. Methods A pair of universal primers were selected in the conserved region of 16S rRNA gene to collect 37 strains of common clinical pathogenic bacteria and extract the bacteria. DOI: 10.3969 / j.issn.NA was detected by PCR and detected by agarose gel electrophoresis Increase the results, at the same time, the sensitivity and specificity of the primers were tested. 106 cases of clinically suspected neonates with sepsis were admitted to hospital within 24 h, under strict aseptic conditions, blood samples were collected and extracted ;; DOI: 10.3969 / j.issn.NA PCR amplification, while blood culture , Blood tests and CRP and ESR levels were detected and compared with 20 non-infected children hospitalized in the same period. Results PCR was used to detect the bacteria. DOI: 10.3969 / j.issn.NA all obtained the expected amplification product of about 371 bp, which only amplified bacteria. DOI: 10.3969 / j.issn.NA, DOI: 10.3969 / j.issn.NA no cross-reaction, the amplification limit of which is 104CFU.L-1 of Escherichia coli. Of the 106 children suspected of sepsis, 15 were positive for blood culture and 36 were positive for PCR. The positive rate of PCR was significantly higher than that of blood (P <0.005). PCR results of 20 non-infected children All negative. According to neonatal sepsis diagnostic criteria, the sensitivity, specificity and diagnostic index of PCR were 82.93%, 96.92% and 179.85, respectively, which was better than blood culture and at least two non-specific indicators of 5 diagnostic methods. PCR method 6 ~ 8 h to detect the results, improved nucleic acid extraction technology can reduce the detection time to 4 ~ 6 h. Conclusion PCR method can detect 16S r bacteria; DOI: 10.3969 / j.issn.NA can quickly determine the presence or absence of bacteria in clinical samples for the early diagnosis of neonatal sepsis has a high sensitivity and specificity.