Detection and genotyping of Borrelia burgdorferi carried by free-living ticks in Arxan,Inner Mongolia,China

doi:10.11853/j.issn.1003.8280.2022.05.006

Abstract

Abstract: Objective To understand the infection and genotypes of Borrelia burgdorferi carried by free-living ticks in Arxan,Inner Mongolia,China,and to provide a basis for local prevention and control of Lyme disease.Methods From May 2020 to May 2021,the drag-flag method was used to collect free-living ticks in Arxan.Tick species were identified by morphology and specific 16S rDNA gene sequencing.Genomic DNA was extracted from 113 randomly selected ticks.Nested-PCR targeting the 5S-23S rRNA intergenic spacer region was used to detect B.burgdorferi,and sequence analysis was performed to determine the genotypes of B.burgdorferi in ticks.The B.burgdorferi carried by 102 Ixodes persulcatus were isolated and cultured in BSK II medium.Multilocus sequence analysis (MLSA) was used to genotype the isolates.Results A total of 295 free-living ticks were collected,which were dominated by I. persulcatus(282/295,95.59%),followed by Dermacentor silvarum(13/295,4.41%).PCR detection of 113 ticks showed that 24 of 100 I. persulcatus carried B. burgdorferi,and the infection rate was 24.00%.Of these,17 were infected with B.garinii(17/24,70.83%),and 7 were infected with B.afzelii (7/24,29.17%).B.burgdorferi was not detected in 13 D.silvarum.Two strains of B. burgdorferi were isolated from the 102 I. persulcatus,which were identified as B. garinii through MLSA.Conclusion The free-living ticks in Arxan in May were dominated by I. persulcatus.The infection rate of B. burgdorferi in I. persulcatus was 24.00%,and the infection was caused by B. garinii and B. afzelii.Two B. garinii strains were isolated from ticks,which suggests that natural infection of spirochetes of Lyme disease was present in Arxan of Inner Mongolia.Local government should pay attention to the risk of B. burgdorferi infection as a causative agent of Lyme disease,monitor the populations,vector ticks,and hosts,and establish a system for the scientific prevention and control of Lyme disease.

Key words: Lyme disease, Tick, Borrelia burgdorferi, Multilocus sequence analysis

摘要： 目的了解内蒙古阿尔山地区游离蜱的伯氏疏螺旋体感染状况及携带的基因型，为当地莱姆病的防控提供依据。方法 2020年5月和2021年5月，在内蒙古阿尔山地区用布旗法采集游离蜱，经形态学和特异性16S rDNA基因检测鉴定蜱种。随机选择113只蜱提取基因组DNA，采用巢式PCR扩增伯氏疏螺旋体5S～23S rRNA基因间隔区，阳性产物测序分析。采用BSK Ⅱ培养基对102只全沟硬蜱中携带的伯氏疏螺旋体进行分离培养，分离出的菌株采用多位点序列分析法（MLSA）鉴定基因型。结果共采获游离蜱295只，经鉴定主要为全沟硬蜱（282/295，95.59%），其次为森林革蜱（13/295，4.41%）。113只蜱的PCR检测结果显示，100只全沟硬蜱中24只携带伯氏疏螺旋体，感染率为24.00%，其中17只感染Borrelia garinii基因型（17/24，70.83%），7只感染B. afzelii基因型（7/24，29.17%），13只森林革蜱中未检出伯氏疏螺旋体。对102只全沟硬蜱中携带的伯氏疏螺旋体进行分离培养，得到2株伯氏疏螺旋体菌株，经MLSA分型鉴定为B. garinii基因型。结论内蒙古阿尔山地区5月游离蜱以全沟硬蜱为主，其中伯氏疏螺旋体感染率为24.00%，感染的基因型为B. garinii和B. afzelii 2种致病基因型，并分离出2株B. garinii菌株，证实内蒙古阿尔山地区存在伯氏疏螺旋体的自然感染。当地应关注伯氏疏螺旋体的感染和致病风险，开展人群、媒介蜱和宿主的监测工作，建立科学合理的莱姆病防控体系。

关键词: 莱姆病, 蜱, 伯氏疏螺旋体, 多位点序列分析

CLC Number:

R384.4
R377

DUAN Li-ke, HOU Xue-xia, ZHANG Lin, BAO Zi-hao, LIU Ze-liang, SHI Qi-yuan, ZHOU Hai-jian, DONG Ai-ying, HAO Qin. Detection and genotyping of Borrelia burgdorferi carried by free-living ticks in Arxan,Inner Mongolia,China[J]. Chinese Journal of Vector Biology and Control, 2022, 33(5): 642-647.

段立科, 侯学霞, 张琳, 包子豪, 刘泽梁, 施淇源, 周海健, 董爱英, 郝琴. 内蒙古阿尔山地区游离蜱携带伯氏疏螺旋体的检测及基因分型研究[J]. 中国媒介生物学及控制杂志, 2022, 33(5): 642-647.

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