Research On The Multiantigen Sequence Typing And The Resistance Genes MtrR And PorB1 Of Neisseria Gonorrhoeae In Taiyuan Shanxi

Objective:1.To study the sensitivity of clinical isolates of Neisseria gonorrhoeae to the seven drugs of ceftriaxone,cefixime,azithromycin,ciprofloxacin,spectinomycin,tetracycline and penicillin in Taiyuan,Shanxi.2.Using the method of the multiantigen sequence typing to explore the genotype of Neisseria gonorrhoeae in Taiyuan,Shanxi Province,and grasping the prevalence of different genotypes of Neisseria gonorrhoeae in this area,and providing theoretical basis for disease prevention and control.3.To analyze the relationship between the sensitivity of cephalosporin in Taiyuan,Shanxi and the resistance genes mtrR and porB1 b.Method:1.From September 2017 to December 2018,61 strains of Neisseria gonorrhoeae in the Second Hospital of Shanxi Medical University of Department of Dermatology,Urology,Drug Sensing,and Shanxi Center for Disease Control and Prevention,were used to detect seven drugs,sputum,azithromycin,ciprofloxacin,spectinomycin,tetracycline,and penicillin,by agar dilution method to study the drug sensitivity.2.The DNA of 61 strains was extracted,and the por and tbpB fragments were amplified and sequenced.The por and tbpB alleles and NG-MAST gene sequences of each strain were analyzed in NG-MAST official website,and the MegaX software was used.The por and tbpB gene sequences were used as phylogenetic trees,and the reliability of the phylogenetic tree was tested by the Bootstrap method to analyze the sequence homology.3.The DNA of the sensitive and resistant strains of ceftriaxone and cefacoxime and one sensitive strain were extracted,and the mtrR and porB1 b gene fragments were amplifiedand sequenced,and the potential mutations were analyzed.Result:1.The resistance rates of ceftriaxone,cefixime,azithromycin,tetracycline,spectinomycin,ciprofloxacin and penicillin were 0,0,4.92%,34.43%,9.8%,100%,100%,respectively.2.(1)After sequencing of 61 strains of Neisseria gonorrhoeae,44 alleles were obtained from por.The genotypes por1132,por2001,por6993,and por581 were the dominant genotypes of por in Shanxi.(2)A total of 26 alleles were obtained from tbpB,among which tbpB4,tbpB186,tbpB1058 and tbpB566 were dominant strains in Shanxi..(3)A total of 49NG-MAST genotypes were found,of which 22 were 21 new strains,and NG-MAST3305 and NG-MAST2083 were popular MAST genotypes in Shanxi.3.(1)The deletion of 33 A in the position of the mtrR promoter was found in 9 strains of low-sensitivity strains.Amino acid substitutions of A39 T,G45D and H105 Y were not completely found in low-sensitivity strains,but not in sensitive strains.(2)The mutations of M18 T,Q143K,and S258 R were present in the amino acid sequence of porB1 b of 9low-sensitivity strains and the sensitive strain,while A121 D,A121G,G120 K was only found in low-sensitivity strains.All strains at positions 215 were replaced.Conclusion:1.Tetracycline,spectinomycin,ciprofloxacin and penicillin are not suitable for the treatment of Neisseria gonorrhoeae in Taiyuan Shanxi,while ceftriaxone,cefixime and azithromycin are still suitable for the treatment of Neisseria gonorrhoeae in Taiyuan,Shanxi.2.(1)The por allele number 90,5692,1123 may be the same source genotype on the short chain.(2)The evolution map of the tbpB of Neisseria gonorrhoeae can be found that the relative relationship of tbpB4,1601,446,75,447,156,and 1058 is relatively close.They may be the same genotypes on the short chain.(3)There are many new genotypes of Neisseria gonorrhoeae in Shanxi,and the genotyping of Neisseria gonorrhoeae in Shanxi requires long-term monitoring.3.(1)The deletion of 33-position A of the mtrR gene,while amino acids the mutations of A39 T,G45D,H105 Y may not be related to the decreased sensitivity of the gonococcal drug;(2)Mutations in the amino acid sequences at positions 120 and 121 of porB1 b mayaffect the drug sensitivity of gonococcal to cephalosporins.