Abstract:We investigated the third-generation cephalosporins-resistant Shigella and its genotype in Ningbo, China, providing a basis for disease prevention and control. Pathogenic bacteria were analyzed by direct isolation combined with enrichment culture isolation. Antimicrobial susceptibility was determined by K-B disk diffusion method and PCR was used for detecting multidrug resistance genes like CTX-M, OXA, TEM and SHV. BLAST analysis was used to determine the genotype. Results showed that 69 strains of third-generation cephalosporins-resistant Shigella were detected by drug sensitivity screening, accounting for 74.19% of ESBLs Shigella. Drug resistance gene CTX-M(CTM-M-1 and CTM-M-9), OXA and TEM were detected. The detection rate were 79.71%, 79.01% and 26.09% respectively.With no CTX-M-2 and SHV, DNA sequence alignment showed CTX-M-1 group were mainly of CTX-M-15 type besides seven other types; CTX-M-9 group were mainly of CTX-M-14 type besides six other types; 49 strains of OXA and 18 strains of TEM were sequenced to be type 1 (OXA-1 and TEM-1 type). The 21 Shigella strains carrying more than two drug resistance genes accounts for 30.43%. Shigella in Ningbo has high third-generation cephalosporins-resistance rate and many kinds of ESBLs enzymes were detected. The mainstream enzyme type was CTX-M,meanwhile they also carried a variety of drug resistance genes, which could bring difficulties to disease prevention and control. The high carrying rate of OXA-1 type suggests that we should pay more attention. The detection rate of group B was higher than that of group D, including not only the phenotype resistance but also the drug-resistance genes; these findings will be useful in the study of the drug resistance prevalence of Shigella.
[1] Mandomando I, Jaintilal D, Pons MJ, et al.Antimicrobial susceptibility and mechanisms of resistance in Shigella and Salmonella isolates from children under five years of age with diarrhea in rural Mozambique[J].Antimicrob Agents Chemother, 2009, 5(6): 2450-2454.doi:10.1128/AAC.01282-08 [2] Nguyen TV, Le PV, Le CH, et al.Antibiotic resistance in diarrheagenic Escherichia coli and Shigella strains isolated from children in Hanoi, Vietnam[J].Antinicrob Agents Chanother, 2005, 49(2): 816-819.doi:10.1128/AAC.49.2.816-819.2005 [3] Tan WB.Advances in genetic mechanism of bacteria resistance[J].J Patho Biol, 2009, 4(7):543-544.(in Chinese) 谭文彬.细菌耐药的基因机制研究进展[J].中国病原生物杂志,2009,4(7):543-544. [4] Ye YW, Wang YS, Shen ZY.National Guide To Clinical Laboratory Procedures[M].The third edition.Nanjing: Southeast University Press, 2003:822-823.(in Chinese) 叶应妩,王毓三,申子喻.全国临床检验操作规程[M].3版.南京:东南大学出版社, 2003: 822-823. [5] Ministry of Health of the PRC.WS 287-2008 The diagnostic criteria of bacterial and amebic dysentery[S].Beijing: People’s Medical Publishing House, 2009.(in Chinese) 中华人民共和国卫生部.WS 287-2008细菌性和阿米巴性痢疾诊断标准[S].北京: 人民卫生出版社, 2009. [6] CLSI.Performance standards for antimicrobial disk susceptibility tests; Approved standard[S].14th ed.Wayne: Clinical and Laboratory Standards Institute, 2014. [7] Clinical and Laboratory Standards Institute.M100-S20 Performance standards for antimicrobial susceptibility testing;twentieth informational supplement[S].Wayne: Clinical and Laboratory Standards Institute, 2014. [8] Ashraf M, Furuta AK, Shimomura K, et al.Genetic characterization of multidrug resistance in Shigella spp.from Japan[J].J Med Microbiol, 2006, 55(pt12): 1685-1691.doi:10.1099/jmm.0.46725-0 [9] Wang Y, Xu YC, Yang QW, et al.The research on TEM-type-β-lactamases among clinical isolates of Escherichia coli and Klebsiella pneumonia in nineteen hospitals[J].J Clinic Lab Sci,2008,26(2):85-89.(in Chinese) 王瑶,徐英春,杨启文,等.19家医院大肠埃希菌和肺炎克雷伯菌中TEM型β-内酰胺酶的研究[J].临床检验杂志,2008,26(2):85-89. [10] Woodfond N.Rapid characterization of beta-lactamases by multiplex PCR[J].Methods Mol Biol, 2010, 64(2): 181-192. [11] Eckert C, GautierV, Saladin-Allard M, et al.Dissemination of CTX-M-type-lactamases among clinical isolates of Enterobacteriaceae in Paris, France[J].Antimicrob Agents Chemother, 2004, 48(4): 1249-1255.doi:10.1007/978-1-60327-279-7_14 [12] Fu YM, Zhang WL, Xu JF, et al.Analysis of CTX-M genotype and resistance of extended-spectrum β-lactamase-producing isolates of Klebsiella pneumonia [J].J Harbin Medical Univ, 2006,40(6):433-436.(in Chinese) 付英梅,张文莉,许建丰,等.产超广谱β-内酰胺酶肺炎克雷伯菌CTX-M基因型和耐药性分析[J].哈尔滨医科大学学报, 2006,40(6):433-436. [13] Zhu DA, Sun JY, Fan HQ, et al.The resistance of Shigella and the genotype screening of extended-spectrum β-lactamase in Shanghai[J].Chin J Infect Chemother, 2009, 9(2): 126-129.(in Chinese) 朱东安,孙景勇,范惠清,等.上海地区志贺菌耐药性及超广谱β-内酰胺酶的基因分型[J].中国感染与化疗杂志,2009,9(2):126-129. [14] Mandomando I, Jaintilal D, Pons MJ, et al.Antimicrobial susceptibility and mechanisms of resistance in Shigella and Salmonella isolates from children under five years of age with diarrhea in rural Mozambique[J].Antimicrob Agents Chemother, 2009, 53(6): 2450-2454.doi:10.1128/AAC.01282-08 [15] Fankhauser C, Zingg W, Francois P, et al.Surveillance of extended spectrum deta lactamase producing Enterobacteriaceae in a Swiss tertiary care hospirtal[J].Swiss Med Wkly, 2009, 139(51-52): 747-751. [16] Bao CM, Guo TS, Cui EB, et al.The research for the distribution of resistant genes of ESBLs-producing isolates of Shigella sonnei [J].Military Med, 2011, 35(2):122-127.(in Chinese) 鲍春梅,郭桐生,崔恩博,等,产ESBLs宋内志贺菌耐药基因分布研究[J].军事医学,2011,35(2):122-127. [17] Yu YS, Ji SJ, Chen YG.Advances of producing CTX-type extended spectrum beta-lactamase[J].Clinic Microbiol Anti-infect Forum, 2004, 10.(in Chinese) 俞云松,季淑娟,陈亚岗.产CTX-M型超广谱β-内酰胺酶研究进展.临床微生物与抗感染论坛, 2004, 10. [18] Nagano Y, Nagano N, Wachino J, et al.Novel chimeric beta-lactamase CTX-M-64, a hybrid of CTX-M-15-Like and CTX-M-14 beta-lactamases, found in a Shigella sonnei strain resistant to various oxyimino-cephalosporins, including ceftazidime[J].Antinicrob Agents Chemother, 2009, 53(1): 69-74.doi:10.1128/AAC.00227-08 [19] Weldhagen G, Poirel L, Nordmann P.Ambler class A extended-spectrum beta-lactamases in Pseudomonas aeruginosa : novel developments and clinical impact[J].Antimicrob Agents Chemother, 2003, 47: 2385-2392 [20] Miao Y, Lu LX, Wu BH, et al.The research on resistant characteristics of OXA-1-type extended-spectrum beta-lactamase-carrying isolates of Shigella in Beijing[J].Hebei Med, 2014, 36(7): 1081-1082.(in Chinese) 苗元,卢立新,吴本和,等.北京地区携带OXA-1型超广谱β-内酰胺酶志贺菌耐药特点研究[J].河北医药,2014,36(7):1081-1082. [21] Ji WJ, Xu XW, Dong F.The research on genotype and resistance of extended-spectrum beta-lactamase-producing isolates of Shigella from pediatric [J].J Microbiol Immunol, 2010, 30(5): 472-476.(in Chinese) 纪文静,徐樨巍,董方.儿科产超广谱 β-内酰胺酶志贺菌的基因型和耐药性研究.中华微生物和免疫学杂志,2010,30(5): 472-476. [22] Zhang CL, Liu QZ, Wang J, et al.Epidemic and virulence characteristic of Shigella spp.with extended-spectrum cephalosporin resistance in Xiaoshan District, Hangzhou, China[J].BMC Infect Dis,2014,14:260.doi:10.1186/1471-2334-14-260 [23] Guo P, He XS, Mei CZ, et al.Detection of Beta lactamase and integron genes in 242 strains of Klebsiella pneumonia isolated in a hospital from 2012 to 2013[J].Chin J Zoonoses, 2015, 31(5): 437-440.(in Chinese) 郭普, 贺晓珊, 梅传忠,等.某医院2012-2013年242株肺炎克雷伯菌β-内酰胺酶及整合子耐药基因检测[J].中国人兽共患病学报2015, 31(5): 437-440.