汉中市中心医院细菌性痢疾志贺菌16S rRNA序列分析、毒力基因与耐药检测

16S rRNA sequence,virulence gene and drug resistance of Shigella dysenteriae in Hanzhong Central Hospital

目的对汉中市中心医院细菌性痢疾患者分离的70株志贺菌进行16S rRNA序列分析、毒力基因与耐药检测。方法对2018年6月至2018年12月我院收治的细菌性痢疾患者进行病原菌分离鉴定,并进行16S rRNA遗传进化分析;PCR法检测其6种毒力基因;K-B法检测其对8种抗生素的敏感性。结果16S rRNA测序结果显示,福氏志贺菌感染率为57.14%(40/70),宋内志贺菌感染率为42.86%(30/70)。16S rRNA系统进化分析结果显示,40株福氏志贺菌与参考菌株(NR026331.1)同源性为95.2%~99.0%,30株宋内志贺菌与参考菌株(NR104826.1)同源性为96.3%~99.9%。毒力基因检测结果表明,70株志贺菌均可以检测出6个相关毒力基因set1A、set1B、sen、Ial、ipaH、virA,毒力基因检出率最高的为ipaH,达到100.00%,其次为Ial、virA,均达到90.00%;其中福氏志贺菌set1A、set1B、Ial基因的检出率高于宋内志贺菌,sen和virA基因的检出率低于宋内志贺菌,二者ipaH基因的检出率都为100.00%。药敏试验结果表明,70株志贺菌耐药率最高的抗生素为氨苄西林,耐药率达到80.00%,其次为强力霉素,耐药率达到75.71%,耐药率最低的为头孢吡肟,耐药率为20.00%;其中40株福氏志贺菌耐药率最高的抗生素为强力霉素,其次为氨苄西林,最低的为头孢吡肟;30株宋内志贺菌耐药率最高的抗生素为氨苄西林,其次为哌拉西林,最低的为氧氟沙星。结论2018年下半年汉中市中心医院从细菌性痢疾患者身上分离得到的志贺菌主要为福氏志贺菌与宋内志贺菌,毒力基因检测和耐药性试验结果显示分离菌株有较强致病性,提示对细菌性痢疾的监测、防控与治疗应进一步加强。

Objective To analyze the 16 S rRNA sequence,virulence genes and drug resistance of 70 Shigella strains isolated from patients with bacillary dysentery in Hanzhong Central Hospital.Methods Isolation and identification of pathogenic bacteria from patients with bacillary dysentery admitted to our hospital from June 2018 to December 2018 and genetic evolution analysis of 16 S rRNA were carried out.The six virulence genes in pathogenic bacteria were detected with PCR.Drug resistance of the pathogen to 8 antibiotics were detected using K-B method.Results The infection rate of Shigella flexneri was 57.14%(40/70)and that of Shigella sonnei was 42.86%(30/70).The 16 S rRNA phylogenetic analysis showed that 40 strains of Shigella flexneri had 95.2%-99.0%homology with the reference strain(NR026331.1),and 30 strains of Shigella sonnei had 96.3%-99.9%homology with the reference strain(NR104826.1).Virulence gene detection showed that the 70 strains of Shigella spp carried 6 virulence genes(set1A,set1B,sen,Ial,ipaH,virA);the detection rate of ipaH was the highest(100.00%),followed by those of Ial and virA(90.00%).The detection rates of set1A,set1B and Ial were higher,while those of sen and virA were lower in Shigella flexneri than in Shigella sonnei,respectively,and the detection rate of ipaH was 100%in both pathogens.Drug sensitivity test showed that the highest resistance rate of the 70 strains of Shigella was to Ampicillin(80.00%),followed by Doxycycline(75.71%),while the lowest resistance rate was to Cefepime(20.00%).Among them,40 strains of Shigella flexneri had the highest resistance rate to Doxycycline,followed by Ampicillin and Cefepime.The highest resistance rate of Shigella sonnei was to Ampicillin,followed by Piperacillin,and that to Ofloxacin was the lowest.Conclusion Shigella flexneri and Shigella sonnei were isolated from bacillary dysentery patients in our hospital in the second half of 2018.The isolated strains were highly pathogenic.The surveillance,prevention,control and treatment of bacillary dysentery should be further strengthened.