标题：PCR-DGGE结合测序在检测混合细菌感染中的价值

      作者：黄君华 1，张书婉 2，张 宁 3
    (1.西安医学院医学技术系，陕西 西安 710021；
2.西安市儿童医院检验科，陕西 西安 710003；
3.西安交通大学医学院第一附属医院检验科，陕西 西安 710061)

      卷次： 2014年25卷8期

      【摘要】 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。

      【关键词】 混合细菌感染；通用引物PCR；变性梯度凝胶电泳；测序；快速检测

      【中图分类号】 R37 【文献标识码】 A 【文章编号】 1003—6350（2014）08—1154—03


Value of PCR-DGGE following by sequencing for detecting mixed bacterial infection.
HUANG Jun-hua 1,
ZHANG Shu-wan 2, ZHANG Ning 3. 1. Department of Medical Technology, Xi'an Medical University, Xi'an 710021,
Shaanxi, CHINA; 2. Clinical Laboratory, Xi'an Children's Hospital, Xi'an 710003, Shaanxi, CHINA; 3. Clinical
Laboratory, the First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an710061, Shaanxi, CHINA

【Abstract】 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.

      【Key words】 Mixed bacterial infection; Broad-spectrum PCR; DGGE; Sequencing; Rapid detection