Molecular typing and antibiotic resistance of Shigella isolated from 2011 to 2014 in Shandong Province
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摘要:
目的 通过对山东省2011-2014年分离的志贺氏菌（Shigella）进行血清、毒力基因、分子分型和药敏等分析，了解山东省志贺氏菌的特性及流行趋势。 方法 用玻片凝集法进行血清分型，PCR方法扩增相关毒力基因，用脉冲场凝胶电泳技术进行分子分型，采用微量肉汤稀释法测定菌株抗生素敏感性。 结果 44株志贺氏菌主要为福氏志贺氏菌（Shigella flexneri）（54.55%）和宋内志贺氏菌（Shigella sonnei）（43.18%）。ipaH、set1、sen、ial毒力基因携带率分别为100%、43.18%、56.82%、50.00%。经脉冲场凝胶电泳分型（pulsed field gel electrophoresis，PFGE），福氏志贺氏菌被分为18种带型，且总体相似度偏低；宋内志贺氏菌被分为14种带型，且89.47%的菌株相似度在90%以上。44株志贺氏菌对15种抗生素中的14种均存在不同程度的耐药。93.18%的菌株为多重耐药菌株。 结论 山东省志贺氏菌以福氏和宋内血清群为主，毒力基因携带率高，有聚类分布出现，耐药严重，应加强对志贺氏菌菌型、溯源和耐药性的监测。 Abstract:Objective To study the characteristics and epidemic trend of Shigella in Shandong province through the analysis of serotype, virulence genes, molecular typing and drug sensitivity. Methods The serotype was classified using the method of slide agglutination. Polymerase chain reaction (PCR) was used to amplify the related virulence genes. The molecular typing was carried out by pulsed field gel electrophoresis (PFGE), and the antibiotic sensitivity of the strains was determined by micro-broth dilution method. Results The main serogroups of 44 Shigella strains were Shigella flexneri (54.55%) and Shigella sonnei (43.18%). The carrying rates of ipaH, Set1, Sen and ial were 100%, 43.18%, 56.82% and 50.00%, respectively. By PFGE typing, the strains of Shigella flexneri were divided into 18 patterns with a low similarity. The strains of Shigella sonnei were divided into 14 patterns, and the similarity of 89.47% of the strains was more than 90%. 44 strains of Shigella had different levels of resistance to 14 of the 15 antibiotics. 93.18% of the strains were multidrug resistant. Conclusion The Shigella in Shandong province is dominated by serogroups of Shigella flexneri and Shigella sonnei, with high virulence gene carrying rate, clustering distribution and severe antibiotic resistance. It is necessary to strengthen the monitoring on serotype, traceability and antibiotic resistance of Shigella in Shandong province. -
Key words:
- Shigella /
- Virulence genes /
- Pulsed field gel electrophoresis /
- Antibiotic resistance
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图 1 福氏志贺氏菌PFGE聚类分析及药敏结果
Figure 1. PFGE Cluster and drug sensitivity analysis of Shigella flexneri
图 2 宋内志贺氏菌PFGE聚类分析及药敏结果
Figure 2. PFGE Cluster and drug sensitivity analysis of Shigella sonnei
表 1 志贺氏菌4种毒力基因引物序列
Table 1. Primer sequences of virulence genes of Shigella
毒力基因 引物序列(5′-3′) 扩增长度(bp) ipaH TGG AAA AAC TCA GTG CCT CT 423 CCA GTC CGT AAA TTC ATT CT set1 TCA CGC TAC CAT CAA AGA 309 TAT CCC CCT TTG GTG GTA sen ATG TGC CTG CTA TTA TTT AT 799 CAT AAT AAT AAG CGG TCA GC ial CTG GAT GGT ATG GTG AGG 320 GGA GGC CAA TTA TTT CC 
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表 2 44株志贺氏菌毒力基因携带情况
Table 2. Virulence genes carrying statistics of 44 Shigella strains
毒力基因型 菌株数 占比(%) ipaH 19 43.18 ipaH+sen 2 4.55 ipaH+set1+sen 1 2.27 ipaH+sen+ial 4 9.09 ipaH+set1+sen+ial 18 40.91
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表 3 44株志贺氏菌对15种抗生素的耐药率
Table 3. Antibiotic resistance rates of 44 Shigella strains to 15 antibiotics
抗生素 敏感(株) 中介(株) 耐药(株) 耐药率(%) 环丙沙星(CIP) 28 0 16 36.36 萘啶酸(NAL) 7 0 37 84.09 氨苄西林(AMP) 1 2 41 93.18 氯霉素(CHL) 4 10 30 68.18 磺胺异恶唑(SMZ) 3 0 41 93.18 复方新诺明(SXT) 6 0 38 86.36 阿莫西林/克拉维酸(AMC) 40 3 1 2.27 四环素(TET) 17 24 3 6.82 多西环素(DOX) 7 5 32 72.73 庆大霉素(GEN) 25 0 19 43.18 头孢曲松(CRO) 30 0 14 31.82 头孢西丁(FOX) 40 2 2 4.55 头孢吡肟(FEP) 34 1 9 20.45 头孢噻肟(CTX) 26 0 18 40.91 亚胺培南(IPM) 44 0 0 0.00
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表 4 44株志贺氏菌多重耐药情况
Table 4. Multidrug resistance rates of 44 Shigella strains
耐药种类数 菌株数 占比(%) 1 2 4.55 2 1 2.27 3 2 4.55 4 1 2.27 5 4 9.09 6 7 15.91 7 10 22.73 8 6 13.64 9 4 9.09 10 6 13.64 11 1 2.27
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