Complete Genome Sequence And Drug Resistance Of Multi-drug Resistant Strain SWU01 Of Klebsiella Pneumoniae

Klebsiella pneumoniae is very common bacteria from clinical specimen,causing typical primary pneumonia,bronchitis and urinary tract infection.In recent years,due to the increased stress of antibiotic selection,the antibiotic abuse problems get more serious,leading to the obvious increase of isolating rate of multidrug-resistant Klebsiella.According to the data released from China Antimicrobial Resistance Surveillance System by the end of 2015,gram-negative bacteria takes up 71.1% of the isolated clinical bacteria,while genus of Klebsiella pneumoniae takes up 19.8% of the isolated gram-negative bacteria.Hence,there is special necessity to conduct profound research on multi drug-resistant Klebsiella.The third generation technique of high-throughout sequencing was conducted on the multi drug-resistant strain SWU01 of Klebsiella for complete genome sequencing.Analysis on the genome sequencing of SWU01 could help us to understand the drug-resistant and spreading mechanism of Klebsiella,which could be of great significance for the prevention and controlling of hospitial infection.Objective: ESBLs,Amp C and carbapenemase together with drug resistance spectrum including 18 antibiotics were analyzed on multi-drug resistant Strain SWU01 of Klebsiella,which could provide scientific references for rational use of antibiotics.High-throughout sequencing technique was used for complete genome sequencing of target strain.Analysis on plasmid of multi drug resistant strains from genetic perspective could unveil the genetic mechanisms for multi drug resistance,which could be helpful for the guidance on prevention and controlling of hospital infection.Method: Cultivate and identify the SWU01 strain of of Klebsiella from one hospital in 2016,to identify its drug resistant phenotype and MIC value of 18anti-microbial(AMK,AMP,SAM,ATM,ETP,NIT,SXT,CIP,TZP,GEN,FEP,CRO,CAZ,CTT,CZO,TOB,IMP and LVX);MLST method was used to determine the type of strain clone;High-throughout sequencing technique was used for complete genome sequencing.Various bio-information analyzing software together with Gene Bank of NCBI were compared with the sequencing,especially for the plasmid p SWU01 with length of 162,552 bp.Information about drug resistant gene were analyzed by bio-information software.Results: 1.SWU01 strain was confired to be Klebsiella.According to the drug sensitive test,it shows resistance rate of 100% to all the 18 anti-microbial,it is positive for ESBLs,Amp C and carbapenemase,proved to be multi drug resistant strain,MLST type is ST11,One set of chromosomal genome was isolated from the strain cultivation.2.The total length of its genome is5,536,506 bp(Gene Bank Accession NO:CP018454-CP018455),average GC content 57.51%,including 6459 protein encoding genes.Based on the analysis of homologous genes,all the homologous genes were linked together to obtain the whole genome level,the relationship with the strain cp006659.2 is definite recently.3.Drug-resistant genes of SWU01 are mainly distributed on plasmid p SWU01.Total length of 162,552 bp,GC content 52.76% p SWU01 harbors250 Open Reading Frames,69 of which are genes relevant to drug-resistance,(including 36 transfer-encoding genes for conjugate plasmid,5 drug-resistance encoding genes(TEM-1?CTX-M-65?SHV-12?rmt B?KPC-2),4 virulence encoding genes,8 transposon(Tn),15 Inserted Sequence(IS)and 1 integrase encoding gene for Type ? Integron.Compared with known drug-resistant plasmids,p SWU01 is 85% identical to the 151,466 bp of drug-resistant plasmid LJ04(Gene Bank Accession NO: KT185451.1),while 76% identical to the 136,848 bp of drug-resistant plasmid pk P1034(Gene Bank Accession NO:KP893385.1);LJ04,p SWU01 and pk P1034 three plasmids carry resistance genes TEM-1,CTX-M-65,SHV-12,rmt B and KPC-2,the expression of multidrug resistance gene sequences are exactly the same,only in their respective plasmid at different relative position.Conclusion: 1.SWU01 strain of of Klebsiella is multi drug resistant,tolerant to all the 18 antibiotics.2.The total length of Klebsiella genome is 5,536,506bp(Gene Bank Accession NO:CP018454-CP018455),average GC content57.51%,including 6459 protein encoding genes,the relationship with the strain cp006659.2 is definite recently;SWU01 contains drug-resistant.Drug-resistant genes are mainly distributed on plasmid p SWU01.3.The total length of p SWU01 is 162,552 bp,GC content 52.76%,including 250 protein encoding genes.harboring rich drug-resistant elements,which is the main reason for the multi-drug resistance.