基于细菌16S rRNA基因扩增临床不常见病原菌的价值

发布时间：2018-05-01 19:39

本文选题：S + rRNA　；参考：《中国感染控制杂志》2016年04期

【摘要】：目的探讨16S rRNA基因扩增与测序在临床不常见病原菌鉴定中的价值,指导临床相关感染的诊治。方法选择临床微生物实验室常规方法难以准确鉴定、无法鉴定或有特殊表型的细菌12株,采用聚合酶链反应(PCR)扩增其16S rRNA基因,测序后进行BLAST比对鉴定菌种,并分析相关感染的临床特点。结果 12株菌经PCR扩增均得到阳性条带(约1 500 bp),均鉴定到种(相似度≥99%),分别为产单核细胞李斯特菌、马耳他布鲁杆菌各2株,死亡梭杆菌、空间罗氏菌、鼻疽奴卡菌、解糖葡萄球菌、放射性根瘤菌、二路普雷沃菌、解甘露醇罗尔斯顿菌及阴道阿托波菌各1株。16S rRNA基因扩增灵敏度高,大肠埃希菌ATCC25922的最低检测限为1.5×101 CFU/mL。12例患者临床资料显示上述菌可引起临床多部位、多类型感染,选用针对性抗菌药物治疗后11例好转,1例死亡。结论 16S rRNA基因测序方法可快速、准确鉴定少见菌、厌氧菌及难培养细菌,为临床不同类型感染的病原学诊断和治疗提供实验室依据。
[Abstract]:Objective to investigate the value of 16s rRNA gene amplification and sequencing in the identification of uncommon clinical pathogens and to guide the diagnosis and treatment of clinically related infections. Methods the 16s rRNA gene was amplified by polymerase chain reaction (PCR) from 12 strains of bacteria which could not be identified or had special phenotype by routine methods in clinical microbiology laboratory. The 16s rRNA gene was amplified by polymerase chain reaction (PCR), and the strains were identified by BLAST comparison after sequencing. The clinical features of related infections were analyzed. Results all the 12 strains were amplified by PCR and positive bands were obtained (about 1 500 BP). All the strains were identified as Listeria monocytogenes, 2 strains of Brucella Malta, 2 strains of Clostridium monocytogenes, 2 strains of Space Roche, and 2 strains of Nucca mallei, respectively. The amplification sensitivity of Staphylococcus glycomyst, Rhizobium radiorrhizobium, Prevow, Rolston and Atobobacter vaginalis were high, and the amplification sensitivity was higher than that of Atobobacter vaginalis. The minimum detection limit of Escherichia coli ATCC25922 was 1.5 脳 101 CFU/mL.12. The clinical data showed that the bacteria could cause multi-site and multi-type infection. Conclusion the 16s rRNA gene sequencing method can quickly and accurately identify rare bacteria, anaerobic bacteria and refractory bacteria, and provide laboratory basis for etiological diagnosis and treatment of different types of infection.
【作者单位】：首都医科大学宣武医院;江西卫生职业学院;
【基金】：首都临床特色应用研究重点专项课题(Z141107002514012)
【分类号】：R440

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