天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (10): 1204-1208.doi: 10.11958/20160440

• 细胞与分子生物学 • 上一篇    下一篇

鮑氏志贺菌临床分离株的毒力基因检测及分子分型

孙阳， 张豪杰， 郭文学， 王哲， 贾宇驰， 祁伟   

  1. 天津医科大学第二医院感染性疾病研究所（邮编 300211）
  • 收稿日期:2016-05-24 修回日期:2016-07-22 出版日期:2016-10-15 发布日期:2016-10-21
  • 通讯作者: 祁伟 E-mail:qiweiwyx@yahoo.com
  • 作者简介:孙阳（1991）， 女， 硕士在读， 主要从事感染性疾病研究

Detection of virulence gene and molecular typing of Shigella boydii isolated from clinical sources

SUN Yang, ZHANG Haojie, GUO Wenxue, WANG Zhe, JIA Yuchi, QI Wei   

  1. Infectious Disease Institute, the Second Hospital of Tianjin Medical University, Tianjin 300211, China
  • Received:2016-05-24 Revised:2016-07-22 Published:2016-10-15 Online:2016-10-21
  • Contact: QI Wei E-mail:qiweiwyx@yahoo.com

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摘要： 目的 了解鲍氏志贺菌临床分离株毒力基因分布、耐药性、分子分型及菌株间流行病学相关性。 方法从我院 2015 年 6 月—10 月间门诊就诊腹泻病患者的粪便中分离收集 9 株鲍氏志贺菌。 采用 K-B 纸片扩散法行抗生素药敏试验， 聚合酶链反应技术检测毒力基因， 脉冲场凝胶电泳技术及多位点序列分型确定分子分型， 分析菌株间流行病学相关性。 结果 9 株鲍氏志贺菌共分为 3 个血清亚型： Ⅰ 型 1 株， Ⅱ 型 3 株， Ⅳ型 5 株。 9 株菌均为多重耐药菌： 对氨苄西林耐药率为 9/9， 对头孢他啶、链霉素、庆大霉素、复方新诺明、头孢噻肟、头孢曲松、四环素、诺氟沙星和左氧氟沙星的耐药率分别为 1/9、4/9、4/9、4/9、5/9、5/9、6/9、6/9、6/9， 未发现阿米卡星、头孢哌酮-舒巴坦和亚胺培南耐药株； ipaH 的检出率为 100%， sen、set1A、set1B、ial、virA、icsA、sigA 的检出率均为 0， pic、sepA、sat 的检出率分别为 4/9、5/9、7/9； 9 株鲍氏志贺菌通过脉冲场凝胶电泳共分为 8 种谱型， 相似性在 63.21%~100%。 多位点序列分析结果显示 6 株为 ST648， 2 株为 ST131、1 株为 ST10。 结论 本院分离的 9 株鲍氏志贺菌共分 3 个血清亚型， 多重耐药情况严重， 菌株间亲缘关系相对较远， 属非暴发病例。

关键词: 志贺菌属, 鲍氏, 微生物敏感性试验, 电泳, 凝胶, 脉冲场, 毒力基因, 脉冲场凝胶电泳, 多位点序列分型

Abstract: Objective To understand genetic distribution, drug resistance, molecular typing and the epidemiological relativeness between strains of the Shigella boydii virulence. Methods Nine Shigella boydii strains were isolated form stool samples of patients with diarrhea from the Enteric Disease Clinic of the Second Hospital of Tianjin Medical University in June - October 2015. The strains were identified by biochemical test and serum agglutination test. Antibiotics susceptibility test was carried out using the Kirby- Bauer method. Polymerase chain reaction was used for detecting virulence genes. Pulsed- field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) technique were used to determine the epidemiological relationship between nine Shigella boydii strains. Results There were three subtypes in nine isolated Shigella boydii samples, including one, three and five isolates in Ⅰ , Ⅱ , Ⅳ subtypes respectively. All of the 9 isolates were multi-drug resistant. The resistant rate of these strains for ampicillin was 100% (9/9), and then the resistant rates of these strains for ceftazidime, streptomycin, gentamicin, trimethoprim/sulfamethoxazole, cefotaxime, ceftriaxone, norfloxacin and levofloxacin were 1/9, 4/9, 4/9, 4/9, 5/9, 5/9, 6/9, 6/9 and 6/9, respectively. All of these strains were sensitive to amikacin, cefperazone-sulbactam and imipenem. The ipaH was carried by all the testing strains, and none of the strains carried the sen, set1A, set1B, ial, virA, icsA and SigA. The detective rates of pic, sepA and sat were 4/9, 5/9 and 7/9 strains, respectively. Nine shigella boydii strains were divided into 8 PFGE types. The similarity between the spectrums of PFGE was 63.21% - 100%. Multilocus sequence typing showed that six isolates were belonged to ST648, two isolates were ST131 and one isolate was ST10. Conclusion Nine isolates of Shigella boydii (divided into three subtyping) isolated from our hospital are multi-drug resistant and they have distant relationships, belonging to the dissemination of case.

Key words: Shigella, Boydii, microbial sensitivity tests, electrophoresis, gel, pulsed-field, virulence genes, pulsedfield gel electrophoresis, multilocus sequence typing