PCR-DGGE结合测序在检测混合细菌感染中的价值

Value of PCR-DGGE following by sequencing for detecting mixed bacterial infection

目的探讨变性梯度凝胶电泳(Denaturing gradient gel electrophoresis,DGGE)结合测序在检测混合细菌感染中的作用,为细菌多重感染的诊断提供新的思路。方法以3份临床培养鉴定为混合感染的胆囊穿刺引流液为对象,提取细菌总DNA并扩增16S rDNA-V3区,后进行DGGE分析,将所得条带切胶纯化再扩增,扩增产物进行测序及BLAST比对,比较比对结果与临床培养结果的一致性。结果 DGGE可以很好地分离16S rDNA-V3区混合PCR产物,每一个扩增子的测序比对结果与临床培养结果一致。结论 PCR-DGGE结合测序的方法可以很好地鉴定临床混合感染标本中的病原菌,可作为快速检测方法在临床应用。

Objective To explore the value of PCR-DGGE and sequencing for detecting mixed bacte- rial infection. Methods A total of 3 samples which has been identified mixed bacterial infection by culture were included. The bacterial DNA was extracted and amplified the 16S rDNA-V3, following by DGGE , se- quencing and BLAST. Then, to compare the consistency of BLAST and cultivation. Results DGGE could differentiate the mixed amplicons of 16SrDNA-V3 sensationally, and the results of BLAST and cultivation were consistent. Conclusion PCR-DGGE and sequencing provide a new way to diagnose mixed bacterial infec- tion, which might be effective for identifying the pathogen in the sample.