Applying high resolution melt analysis to discriminate VEB-3 hypotype of the clinical gram negative isolates

VernacularTitle:建立高分辨率熔解曲线法筛选含VEB-3型基因菌株
Author: Gang LI ; Quhao WEI ; Yingqiao NI ; Yanyan WANG ; Xin DU ; Xiaofei JIANG
Publication Type:Journal Article
Keywords: VEB-3 hopotype; High resolution melt; Extended spectrum β-lactamase
From: Chinese Journal of Microbiology and Immunology 2010;30(10):956-958
CountryChina
Language:Chinese
Abstract: 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.