中国比较医学杂志2017，Vol.27Issue(5)：94-98,5.DOI:10.3969.j.issn.1671-7856.2017.05.020
金黄色葡萄球菌、绿脓杆菌和肺炎克雷伯杆菌多重PCR方法的建立与初步应用
Establishment and application of a multiplex PCR method in Staphylococcus aureus、Pseudomonas aeruginosa and Klebsiella pneumoniae
摘要
Abstract
Objective Aiming at detecting Staphylococcus aureus、Pseudomonas aeruginosa and Klebsiella pneumoniae in laboratory animals,the paper provides a rapid,sensitive and simple test method.Methods According to Staphylococcus aureus nuc gene,Pseudomonas aeruginosa LasI gene,Klebsiella pneumonia PhoE gene and general 16S rRNA gene, designed specific primers;Through the optimization of multiplex PCR primer concentrations and annealing temperature, the specificity and sensitivity of detection, establishing multiplex PCR system.Application of the PCR system test specimens of artificial infections and experiment animal feces is compared with traditional test method.Results Multiplex PCR amplification of Staphylococcus aureus (153 bp), Pseudomonas aeruginosa (600 bp) with Klebsiella pneumoniae (368 bp) and general (520 bp).The multiplex sensitivity for the purpose of 10pg, specificity of detection was not detected from other pathogens.Application of establishing multiplex PCR system to detect the artificial positive samples, and detect 1 Pseudomonas aeruginosa positive case in 76 fecals.Conclusions This paper established the multiplex PCR method which has the advantages of specific,sensitive,simple and rapid, and provides a reliable way for rapid test in laboratory animals microbiology.关键词
金黄色葡萄球菌/绿脓杆菌/肺炎克雷伯杆菌/多重PCRKey words
Staphylococcus aureus/Pseudomonas aeruginosa/Klebsiella pneumoniae/Multiplex PCR分类
医药卫生引用本文复制引用
祝岩波,王俊霞,郑龙,尤红煜,王璇,梁晓亮,刘健敏,张东明,连伟光,栗彦宁..金黄色葡萄球菌、绿脓杆菌和肺炎克雷伯杆菌多重PCR方法的建立与初步应用[J].中国比较医学杂志,2017,27(5):94-98,5.基金项目
国家科技支撑计划(2015BAI07B02-05). （2015BAI07B02-05）
