艰难梭菌菌型中鞭毛蛋白基因 fliC 和 fliD 的运动性和基因型多样性。
Motility and the genotype diversity of the flagellin genes fliC and fliD among Clostridioides difficile ribotypes.
机构信息
Department of Medical Microbiology, Medical University of Warsaw, Warsaw, Poland.
Department of Medical Microbiology, Medical University of Warsaw, Warsaw, Poland; The member of European Society of Clinical Microbiology and Infectious Disease (ESCMID) Study group for Clostridioides difficile (ESGCD), Poland.
出版信息
Anaerobe. 2022 Feb;73:102476. doi: 10.1016/j.anaerobe.2021.102476. Epub 2021 Nov 13.
OBJECTIVE
The motility and genotype of the flagellin fliC and fliD genes were investigated in 82 Clostridioides difficile isolates belonging to the ribotypes (RTs): 027 (n = 41), 176 (n = 17), 023 (n = 8), 017 (n = 6) and 046 (n = 10). The reference C. difficile strains 630 and M120 were included as controls for the motility assay.
METHODS
A Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) was used to exclude the genetic relatedness of C. difficile isolates belonging to the same RT. The variability of the fliC and fliD genes was determined by PCR-restriction fragment length polymorphism (RFLP) analysis and Sanger sequencing. The motility assay was carried out with 0.175% BHI agar tubes and BHI solid media plates with 0.4% agar.
RESULTS
The highest motility was observed in C. difficile RT023 isolates (p < 0.01), followed by RTs 027 and 176. C. difficile isolates of RTs 017 and 046 were less motile than RTs 027, 176 and 023 (p < 0.01). The fliC and fliD genes were present in all clinical isolates irrespective of the motility results. In the fliC gene analysis, four different RFLP groups were identified (I, II, VII, X). The fliC group VII was identified in two RTs (027 and 176), whereas the remaining three groups (I, II and X) belonged to a single RT 046, 017 and 023, respectively. The fliD gene analysis identified four new RFLP groups (a, b, c and d).
CONCLUSIONS
C. difficile RT023 is highly motile and its motility is comparable to the hypervirulent RT027 and its genetic relative RT176.
目的
研究了 82 株艰难梭菌分离株的 flagellin fliC 和 fliD 基因的运动性和基因型，这些分离株属于核糖体分型（RTs）：027（n=41）、176（n=17）、023（n=8）、017（n=6）和 046（n=10）。参考菌株 630 和 M120 艰难梭菌被纳入运动性检测的对照组。
方法
采用多位点可变串联重复分析（MLVA）排除属于同一 RT 的艰难梭菌分离株的遗传相关性。通过 PCR-限制性片段长度多态性（RFLP）分析和 Sanger 测序确定 fliC 和 fliD 基因的变异性。运动性检测使用 0.175% BHI 琼脂管和含有 0.4%琼脂的 BHI 固体培养基板进行。
结果
观察到 C. 艰难梭菌 RT023 分离株的最高运动性（p<0.01），其次是 RT027 和 RT176。RT017 和 RT046 的 C. 艰难梭菌分离株的运动性低于 RT027、176 和 023（p<0.01）。无论运动性结果如何，所有临床分离株均存在 fliC 和 fliD 基因。在 fliC 基因分析中，确定了四个不同的 RFLP 组（I、II、VII、X）。fliC 组 VII 存在于两个 RT（027 和 176）中，而其余三个组（I、II 和 X）分别属于单个 RT 046、017 和 023。fliD 基因分析确定了四个新的 RFLP 组（a、b、c 和 d）。
结论
C. 艰难梭菌 RT023 高度运动，其运动性与高毒力 RT027 及其遗传相关 RT176 相当。
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