Application of 16S rDNA full-length high-throughput sequencing in the study of tick-borne pathogen biodiversity

doi:10.11853/j.issn.1003.8280.2021.04.004

Abstract

Abstract: Objective To study the characteristics of tick-borne pathogenic populations using 16S rDNA full-length sequencing and laboratory test data, and to provide a basis for pathogen control. Methods Fifty ticks were selected from the dominant tick species in Qi county, Shanxi province, China. Whole genome DNA was extracted after morphological and genetic identification. The species and taxonomic abundance of microorganisms in ticks were analyzed using 16S rDNA full-length high-throughput sequencing on the PacBio platform. According to the 16S rDNA sequence analysis, the specific genes of tick-borne pathogens were identified by PCR for a comprehensive analysis of tick-borne pathogens. Results The 16S rDNA full-length sequencing analysis showed that all the 50 ticks carried Coxiella burnetii and Pseudomonas, 8 ticks carried Anaplasma phagocytophilum, some ticks carried Rickettsia and Ehrlichia (only identified at the genus level), and Borrelia was not detected. The PCR results showed that C. endosymbiont, Borrelia burgdorferi, B. miyamotoi, and spotted fever group rickettsia were detected in 50 ticks. Conclusion High-throughput 16S rDNA full-length sequencing can simultaneously detect a variety of pathogens and identify the main tick-borne pathogens in the area, though some pathogens are identified at the genus level, which requires combination with pathogen-specific tests for comprehensive detection of tick-borne pathogens in the area.

Key words: 16S rDNA full-length sequencing, Laboratory molecular detection, Tick-borne pathogens, Biodiversity

摘要： 目的利用16S核糖体DNA（16S rDNA）全长测序方法，结合实验室检测数据，研究蜱中病原微生物种群特点，为病原体控制提供依据。方法选取山西省祁县优势蜱种50只，经形态学和基因鉴定后，提取全基因组DNA，采用PacBio平台、高通量16S rDNA全长测序方法，分析蜱中微生物种类及各种群丰度情况；根据16S rDNA全长序列分析结果，利用PCR检测方法对可能存在的蜱媒病原体特异性基因进行鉴定，较为全面地分析蜱媒病原体携带情况。结果 16S rDNA全长测序分析表明50只血蜱均携带贝氏柯克斯体、假单胞菌，8只血蜱携带嗜吞噬细胞无形体，有部分血蜱携带立克次体、埃立克体，但仅匹配到属一级的分类信息，无法确定所携带的基因种，在50只血蜱中未检出疏螺旋体。PCR检测结果表明，50只血蜱中检测到柯克斯体内共生体、伯氏疏螺旋体、米氏疏螺旋体和斑点热群立克次体。结论高通量16S rDNA全长测序分析可以同时检测出多种病原体，可提示当地的主要蜱媒致病菌，但也存在检测不全面、无法匹配到物种的缺点，与病原体特异检测方法相结合，能更全面地了解当地血蜱携带病原体的状况。

关键词: 16S核糖体DNA全长测序, 实验室分子检测, 蜱媒致病菌, 生物多样性

CLC Number:

R384.4
R372

YANG Xiao-na, ZHANG Lin, HOU Xue-xia, HAO Qin. Application of 16S rDNA full-length high-throughput sequencing in the study of tick-borne pathogen biodiversity[J]. Chines Journal of Vector Biology and Control, 2021, 32(4): 404-411.

杨小娜, 张琳, 侯学霞, 郝琴. 16S rDNA全长高通量测序在蜱媒病原生物多样性研究中的应用[J]. 中国媒介生物学及控制杂志, 2021, 32(4): 404-411.

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