聚合酶链反应检测革兰阴性细菌16SrRNA

16S rRNA Gene Detection of Gram-negative Bacteria by PCR

【目的】寻找诊断细菌感染病原更为敏感和有效的方法。【方法】根据细菌１６ＳｒＲＮＡ基因的高度保守性，设计合成革兰阴性细菌的共同引物，采用聚合酶链反应检测已知的革兰阴性细菌９株，革兰阳性菌４株。【结果】革兰阴性菌检测阳性率为１００％，（检测革兰阳性细菌４株结果均为阴性）。采用倍比稀释法检出大肠杆菌的最低浓度为４ＣＦＵ／ｍｌ。【结论】所用引物具有良好的特异性和敏感性，能达到初步鉴定细菌种属的目的。

Objective To find out a sensitive and effective method for diagnosis of pathogens in bacterial infection. Methods According to the highly conservative nature of bacteria 16S rRNA gene,a set of primers with broad range for Gram-negative bacteria was designed.Nine known strains of Gram-negative bacteria were detected by polymerase chain reaction(PCR). Results All 9 strains of Gram-negative bacteria were positive,while 4 strains of Gram-positive were negative.The lowest concentration of Escherichia coli detected by serial dilution was 4 CFU/ml. Conclusions This set of PCR primers with broad range has high specificity and sensitivity,achieving a preliminary identification of bacteria species.