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Evaluation of four molecular typing methods in the homology analysis of Acinetobacter species

LIU Wenjing;ZHANG Xiaojiang;YANG Qiwen;XU Yingchun;Department of Clinical Laboratory,Chinese Academy of Medical Sciences/Peking Union Medical College/Peking Union Medical College Hospital;  
Objective To evaluate four molecular typing methods(PFGE,RAPD,Diversilab and MLST)in the homology analysis of Acinetobacter species.Methods Carbapenem-resistant mutant strains of Acinetobacter were screened by imipenem and meropenem in vitro.Molecular identification was carried out by using amplifed ribosomal DNA restriction analysis(ARDRA).The homology of parental sensitive strains and resistant mutant strains was analyzed by using four typing techniques:pulsed field gel electrophoresis(PFGE),random amplification of polymorphic DNA(RAPD),automatic Rep-PCR(Diversilab)and multilocus sequence typing(MLST).Results ARDRA of carbapenems parental susceptible strains and resistant mutant strains were consistent,including Acinetobacter genospecies 3,Acinetobacter baumannii genospecies 2 and Acinetobacter genospecies 13 TU.The strains were typed by PFGE,RAPD,Diversilab and MLST.It was found that the homology between drug-resistant mutants selected in vitro and parental susceptible strains is closely related.RAPD and Diversilab had poorer resolution than PFGE,and similar strains could not be distinguished.PFGE showed that there were more than 7 bands between the parental susceptible strains of Pu53 and their drug-resistant mutants,and the parental susceptible strains of pu61 and their drug-resistant mutants,and there was no relationship between them.RAPD and Diversilab DNA are closely related.Compared with other typing methods,MLST showed that strains with the same genetic background had the same ST type,although gpi-PCR and gdhB sequencing or amplification of Acinetobacter genospecies 3 failed.RpoD non-degenerate primers did not amplify Acinetobacter genome 13 TU and Acinetobacter genospecies 3.Conclusion RAPD and Diversilab were suitable for rapid homology analysis in laboratory,PFGE and MLST should be selected if there were not well differentiated results.
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