Vilnius City Clinical Hospital, Vilnius, Lithuania.
Department of Microbiology and Biotechnology, Institute of Biosciences, Life Sciences Center, Vilnius University, Vilnius, Lithuania.
Eur J Clin Microbiol Infect Dis. 2019 Oct;38(10):1953-1959. doi: 10.1007/s10096-019-03633-9. Epub 2019 Jul 20.
Clostridioides difficile has become the leading nosocomial Gram-positive pathogen in the developed countries. In Lithuania, the national surveillance program for C. difficile started in 2017. Enzyme immunoassay, the real-time PCR system, and culture are used for laboratory confirmation of C. difficile infection in Lithuanian clinical laboratories. No reference laboratory for C. difficile is present in Lithuania. Fifty-eight isolates of C. difficile were collected in 2016 and 2017 in two hospitals using real-time PCR and culture methods. Agarose gel-based PCR ribotyping, multilocus variable number tandem repeats analysis (MLVA), and multilocus sequence typing (MLST) were used for the genotypic characterization of 28 isolates. PCR ribotyping and MLST showed that 78.6% of the tested toxigenic isolates belong to the ribotype RT027/ST1. Using MLVA, 95.5% of RT027 isolates were genetically related. MLVA revealed three clonal complexes in RT027. Six non-RT027 isolates showed four different electrophoretic profiles in PCR ribotyping and were assigned to the MLST sequence types ST2, ST13, ST54, and ST63. The highest discriminatory power showed the genotyping by MLVA. In total, 20 MLVA profiles were identified. This genotyping technique allowed to identify four groups of RT027/ST1 isolates that were indistinguishable by PCR ribotyping and MLST. Our study is the first genotypic characterization of C. difficile isolates in Lithuania. We observed a high prevalence of presumptive RT027 that suggests unfavorable epidemiological situation in Lithuania. Our results stress for implementation of genotyping of C. difficile isolates in Lithuanian surveillance.
艰难梭菌已成为发达国家中主要的医院内革兰阳性病原体。在立陶宛，艰难梭菌的国家监测计划于 2017 年开始。酶免疫分析、实时 PCR 系统和培养用于立陶宛临床实验室中艰难梭菌感染的实验室确认。立陶宛没有艰难梭菌的参考实验室。2016 年和 2017 年，使用实时 PCR 和培养方法在两家医院共收集了 58 株艰难梭菌。琼脂糖凝胶 PCR 核糖体分型、多位点可变数目串联重复分析（MLVA）和多位点序列分型（MLST）用于 28 株分离株的基因特征描述。PCR 核糖体分型和 MLST 显示，78.6%的测试产毒分离株属于 RT027/ST1 核糖体型。使用 MLVA，95.5%的 RT027 分离株在遗传上相关。MLVA 揭示了 RT027 中的三个克隆复合体。6 株非 RT027 分离株在 PCR 核糖体分型中显示出四种不同的电泳图谱，被分配到 ST2、ST13、ST54 和 ST63 型 MLST 序列类型。MLVA 显示出最高的分辨力。共鉴定出 20 种 MLVA 图谱。这种基因分型技术能够鉴定出 RT027/ST1 分离株的四个组，这些组在 PCR 核糖体分型和 MLST 中无法区分。本研究是立陶宛艰难梭菌分离株的首次基因特征描述。我们观察到假定的 RT027 高流行率，这表明立陶宛的流行病学情况不利。我们的结果强调了在立陶宛监测中实施艰难梭菌分离株基因分型的必要性。
