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Experimental Research Of Bacteria Identification And Homology Analysis By Use Of Matrix-assisted Laser Analysis/Ionization Flight Time Mass Spectrometry

Posted on:2019-07-14Degree:MasterType:Thesis
Country:ChinaCandidate:J M XuFull Text:PDF
GTID:2394330542997307Subject:Clinical Laboratory Science
Abstract/Summary:PDF Full Text Request
Objective:Toevaluatethefeasibilityofmatrix-assistedlaser desorption/ionization-time of flight mass spectrometry?MALDI-TOF MS?in bacteria identification and drug-resistant bacteria's homology analysis in clinical laboratory.Methods:Four hundred and forty strains of non-repeated samples were obtained in our hospital from January 2015 to January 2016.Firstly,we took advantage of VITEK 2 automatic microbiological identification&drug sensitive system?traditional biochemical reaction technique?and VITEK MS automatic rapid microbiological mass spectrometer?MALDI-TOF MS technique?to identify these strains,respectively.Then,the inconsistent results were re-confirmed with the ribosomal 16S rRNA gene sequence analysis.Concurrently,to evaluate the advantages of MALDI-TOF MS in turn-around time?TAT?,we collected 95 strains of Klebsiella pneumoniae screened by modified Hodge test?MHT?.By use of multilocus sequence typing?MLST?and Microflex TMM mass spectrometer?MALDI-TOF MS technique?,we analyzed bacteria's homology of the 95 strains respectively.Results:Using the result of VITEK 2 as the reference,the total accuracy rate of the 440 strains identified by VITEK MS was 96.6%at species level,and 99.1%at genus level.The results of 15 inconsistent strains identified by 16S rRNA showed that there were 3 different identification at species level by VITEK MS,and there were 12different identification at species level by VITEK 2?P=0.034?.However,it took nearly8 hours per plant for VITEK 2 comparing 8 hours for VITEK MS to complete the tests.and it took only 1 min per strain.The results of 95 CRKP MLST typing were divided into 9 sequence types,ST11 accounted for 91.6%.The results of cluster analysis obtained by MicroflexTM mass spectrometer were divided into 2 types,5 subtypes.The?b and?c had a close relationship,and?a and?b??c had a distant relationship.Conclusion:The above results indicate that MALDI-TOF MS could be applied to identify common bacteria from the clinical departments.The identification ability of MS has more advantageous than traditional biochemical reaction identification.In TAT,MALDI-TOF MS greatly shortened identification time..According to the MLST classification of CRKP,there are 9 sequence types in 95 strains,and the main type of the classification is ST11,which is consistent with the domestic popular investigation report.The results of the MALDI-TOF MS cluster analysis showed that the correlation analysis of 95 strains was more detailed than that of MLST.
Keywords/Search Tags:Matrix-assisted laser Desorption/ionization-time of flight mass spectrometry, 16S rRNA, Identity, Turn-around time, Multilocus sequence typing
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