16SrRNA基因聚合酶链反应快速检测烧伤脓毒症细菌病原体的研究

目的为早期诊断烧伤脓毒症，探索一种能迅速检测细菌感染的方法。方法按标准收集可疑脓毒症患者39人外周静脉血标本共41份。将每份标本一分为二分别行血培养和用细菌通用引物行16SrRNA基因聚合酶链反应检测，用琼脂糖凝胶电泳分析PCR产物，比较两种方法在检测细菌病原体时的快速性和敏感性，分析血培养及药敏结果。结果16SrRNA基因PCR方法的阳性率（41．16％）是血培养阳性率（21．95％）的近两倍。P〈0．05。16SrRANA基因PCR方法出结果需4、5h，血培养需12～24h（多数需2、3d）。9例血培养阳性患者中，8例为单一铜绿假单孢菌感染，1例为单一溶血链球菌感染。结论16SrRNA基因PCR检测细菌感染的方法具有特异性、快速性、敏感性的优点。在该中心，使用抗生素治疗后，引起脓毒症的细菌主要为铜绿假单孢菌。该菌对目前常用广谱抗生素耐药。

Study on rapid detection of bacterial in burn sepsis by16SrRNA genes polymerase chain reaction

Objective] To explore a method for rapid detection of bacterial infection in clinic to diagnose burn sepsis early. Methods] 41 blood samples of 39 cases with suspect burn sepsis were collected in the department of burn according to some criteria. Every blood sample was divided into two parts. One was sent for blood culture (BC), the other was detected by way of 16SrRNA gene Polymerase Chain Reaction(PCR) with universal primers of bacterial and PCR products were analyzed by electrophoresis using agarose gel. Then analyzed outcomes of two parts and compared the rapidity and sensitivity of two methods to detect bacterial of patients with burn sepsis. Analyed the outcomes of BC and the tests of drug sensitivity. Results] The positive rate of the way of 16SrRNA gene PCR (41.16%) is nearly two times of the way of BC(21.95%). The outcome has statistical significance according to chi square test of paired sample, (P <0.05). Meantime the spent time on the way of 16SrRNA gene PCR is shorter than that on the way of BC. The former is 4, 5 hours while the latter is 12~24 hours(mainly 2~3 days). 8 of 9 positive samples with blood culture were the infections of single Pseudomonas aeruginosa and only one was the infection of single streptococcus hemolyticus. Pseudomonas aeruginosa resists the most broad-spectrum antibiotics that are used presently in our hospital from the tests of drug sensitivity. Conclusions] The method of 16SrRNA gene PCR in detecting bacterial has the advantage of specificity, rapidity and sensitivity. The main bacterial that leads to burn sepsis is the single Pseudomonas aeruginosa after using antibiotics in the department of burn of our hospital. Pseudomonas aeruginosa resists the most broad-spectrum antibiotics, which are used presently in our hospital from the tests of drug sensitivity.