摘要
目的将焦磷酸测序技术应用于败血症常见病原体的检测鉴定。方法根据细菌16S rRNA序列的特点,设计焦磷酸测序用的PCR扩增引物和测序引物,提取败血症常见病原体的基因组DNA,进行焦磷酸测序,分析获得的序列,进行比对,对病原体种类作出判断。结果经焦磷酸测序可以获取有效长度28 bp的序列,使用该测序序列可以有效区分化脓性链球菌、肺炎链球菌、大肠埃希菌、铜绿假单胞菌、肺炎克雷白杆菌、脑膜炎双球菌、沙门氏菌,但不能区分金黄色葡萄球菌和表皮葡萄球菌。结论应用焦磷酸测序可以较有效地对败血症常见病原体的种类实现种属水平的区分。
Objective To apply pyrosequencing technique in the detection of the common pathogens in sepsis. Methods The primers for amplification and sequencing in pyrosequencing were designed according to alignment of the bacterial 16S rRNA sequence. Bacterial genomic DNA was extracted for pyrosequencing, and the pathogen species were determined according to the sequencing data obtained. Results Pyrosequencing effectively yielded the sequencing data of the 28 bp sequences of the pathogens and clearly distinguished the pathogen species of Streptococcus pyogenes, Streptococcus pneumonia, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Neisseria meningitides, and Salmonella, but failed to distinguish Staphylococcus epidermidis from Staphylococcus aureus. Conclusion Pyrosequencing technique can effectively distinguish the common pathogens in sepsis at the species level.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2013年第7期1053-1056,共4页
Journal of Southern Medical University
基金
广东省医学科研基金(A2012358)
南方医院院长基金(2009C020)
