CAJ | 학술논문

目的建立高分辨率熔解曲线法筛选含VEB-3型基因菌株.方法不重复收集2003年1-12月革兰阴性杆菌产超广谱β-内酰胺酶(ESBL)菌株292株,用酚-氯仿法抽提基因组DNA;首先以基因组DNA为模板,PCR扩增VEB基因;挑选阳性菌株34株再PCR扩增包含168位点的一段约110 bp的基因片段,高分辨率熔解曲线(high resolution melt,HRM)分析后测序验证.结果 VEB-3亚型与其他亚型基因通过HRM分析具有显著的差别;测序结果与HRM分型结果一致.结论应用HRM技术可以准确快速筛选含VEB-3型基因菌株.
목적 건립고분변솔용해곡선법사선함VEB-3형기인균주.방법 불중복수집2003년1-12월혁란음성간균산초엄보β-내선알매(ESBL)균주292주,용분-록방법추제기인조DNA;수선이기인조DNA위모판,PCR확증VEB기인;도선양성균주34주재PCR확증포함168위점적일단약110 bp적기인편단,고분변솔용해곡선(high resolution melt,HRM)분석후측서험증.결과 VEB-3아형여기타아형기인통과HRM분석구유현저적차별;측서결과여HRM분형결과일치.결론 응용HRM기술가이준학쾌속사선함VEB-3형기인균주.
Objective To establish a new method, applying high resolution melt, to discriminate the VEB-3 hypotype from the clinical gram negative isolates. Methods From January to December 2003,292 consecutive and non-repetitive gram-negative bacteria producing VEB extended spectrum β-lactamase (ESBL) were collected. Extract the DNA of clinical gram negative isolates with phenol-chloroform. PCR was performed to amplify the VEB gene with the DNA being template. After that, we amplify the fragment of VEB gene containing the position 168. Then we detect the high resolution melt curve and analyze them. At last, we analyze the results of sequence and high resolution melt( HRM ). Results VEB-1 and VEB-3 gene are markedly different through HRM analysis. Conclusion It is accurately and quickly for us to identify the VEB-3 from other hypotype through the technology of HRM.