Modern Features of Pathogenic Leptospira Isolation and Identification in Siberia and the Far East

N. V. Breneva, E. Y. Kiseleva, M. B. Sharakshanov, S. Borisov, S. E. Budaeva, S. Balakhonov
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Abstract

Recently, pathogenic Leptospira culture isolation is an extremely rare phenomenon in Russia.The aim of our work was to synthesize the lessons learned at the Irkutsk Anti-Plague Institute from Leptospira culture isolation and identification since 2011.Materials and methods. Material from eight individuals with suspected leptospirosis and from 942 small mammals (SM) was examined using PCR and microscopic agglutination test (MAT), from humans and 260 SM, applying bacteriological method. Bacteriological temperature test, MAT, PCR, MLST and MALDI-TOF mass spectrometry with the original Leptospira protein profiles base were used to identify cultures. Six complete genomes were generated at the Central Research Institute of Epidemiology of the Rospotrebnadzor.Results and discussion. Leptospira have not been isolated from humans against the background of taking antibiotics, despite the positive PCR and MAT results. Four cultures of Leptospira borgpetersenii of the Javanica serogroup and three L. kirschneri (Grippotyphosa) have been isolated from SM. The results of identification using MLST scheme No. 1 and MALDI-TOF MS are identical. MLST in silico has shown the uniformity of two Grippotyphosa serogroup strains from Primorie and Khabarovsk with a sequence-type (ST) profile 110:100:94. ST146 is determined in four Javanica serogroup strains according to scheme No. 1, and unique single nucleotide polymorphisms are detected according to schemes No. 2–3. Thus, in Siberia and the Far East, between 2012 and 2016, seven pathogenic Leptospira cultures were isolated from carriers in natural foci; carrier infection rate being12.0–48.9 %. Javanica serogroup strains differ in the MLST profile characteristics and adapt to nutrient media for a longer time than Grippotyphosa serogroup strains.
西伯利亚和远东地区病原性钩端螺旋体分离和鉴定的现代特征
我们的工作旨在总结伊尔库茨克反鼠疫研究所自2011年以来在钩端螺旋体培养分离和鉴定方面的经验教训。使用 PCR 和显微凝集试验(MAT)对 8 名疑似钩端螺旋体病患者和 942 只小型哺乳动物(SM)的材料进行了检测,并使用细菌学方法对人类和 260 只小型哺乳动物的材料进行了检测。细菌学温度测试、MAT、PCR、MLST 和 MALDI-TOF 质谱法与原始钩端螺旋体蛋白图谱基础一起用于鉴定培养物。Rospotrebnadzor 流行病学中央研究所生成了六个完整的基因组。尽管 PCR 和 MAT 检测结果呈阳性,但在服用抗生素的情况下,人类尚未分离出钩端螺旋体。从 SM 中分离出 4 株 Javanica 血清群的 Leptospira borgpetersenii 和 3 株 L. kirschneri(Grippotyphosa)。使用 MLST 方案 1 和 MALDI-TOF MS 进行鉴定的结果相同。MLST in silico 显示,来自 Primorie 和哈巴罗夫斯克的两株 Grippotyphosa 血清群菌株的序列类型(ST)特征为 110:100:94,具有一致性。根据第 1 号方案,确定了四个爪哇血清群菌株的 ST146,根据第 2-3 号方案,检测到了独特的单核苷酸多态性。因此,2012年至2016年期间,在西伯利亚和远东地区的自然病灶中,从带菌者身上分离出了7株致病性钩端螺旋体培养物;带菌者感染率为12.0%-48.9%。与Grippotyphosa血清群菌株相比,Javanica血清群菌株在MLST谱特征上有所不同,适应营养培养基的时间更长。
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