黑龙江口岸蜱媒病原的多样性及其复合感染调查

发布时间：2018-10-18 20:40

【摘要】：目的:为了研究黑龙江地区密山、绥芬河、嘉荫、东宁、虎林、同江、逊克、萝北、牡丹江、黑河、饶河11口岸蜱类分布、携带及复合感染新发蜱传病原体的情况。方法:2014年4月到2015年10月,综合黑龙江口岸森林植被和蜱媒病的生态学、生物学历史资料,选择密山、绥芬河、嘉荫、东宁、虎林、同江、逊克、萝北、牡丹江、黑河、饶河11口岸作为调查样点,利用布旗法采集样本蜱,经形态学鉴定后,根据地区及种类进行编号,单只检测。蜱独立分装,在管壁上做好编号,放入无菌钢珠,在低温环境下利用研磨仪将蜱研磨至粉末状,4℃保存,按照试剂盒操作步骤提取蜱组织DNA,通过建立五种新发蜱传疾病病原体(伯氏疏螺旋体、斑点热群立克次体、巴尔通体、巴贝西原虫、嗜吞噬细胞无形体)特异性引物,利用PCR或巢式PCR方法对所有样本进行特异性片段扩增,选取PCR阳性片段送测序公司进行测序,所获得的序列与网上(NCBI)序列进行同源性比较,并做进化树分析,以确定并分析病原体在媒介蜱中的地理及特征分布。结果:1、蜱类构成共采集1306只蜱标本,经分类鉴定分属于革蜱属的森林革蜱、硬蜱属的全沟硬蜱和血蜱属的嗜群血蜱和日本血蜱。其中森林革蜱Dermacentor silvarum、625只占总数的47.86%,为优势种群,全沟硬蜱Ixodes persulcatus332只占25.42%,嗜群血蜱Haemaphysalis concinna 308只占23.58%,日本血蜱Haemaphysalis japonica 41只占3.14%。2、病原检测结果本研究在所检测的1306份样本中,共检测出1例巴贝西原虫阳性样本,528例斑点热群立克次体阳性样本、11例巴尔通体阳性样本、31例嗜吞噬细胞无形体阳性样本、11例莱姆病螺旋体阳性样本,感染率分别为0.8‰、404.3‰、8.4‰、23.7‰和8.4‰。对5种新发蜱媒疾病的病原体进行检测,共发现8种病原体,分别是嘎氏疏螺旋体Borrelia afzelii、伽氏疏螺旋体Borrelia garinii、巴贝西原虫Babesia spp.、劳氏立克次体Rickettsia raoultii、新塔拉塞威奇立克次体Candidatus Rickettsia tarasevichiae、黑龙江立克次体Rickettsia heilongjiangensis、巴尔通体Bartonella spp.、嗜吞噬细胞无形体Anaplasma phagocytophilum。本研究结果显示,黑龙江11个口岸地区至少存在两种莱姆病螺旋体(Borrelia garinii、Borrelia afzelii),三种斑点热群立克次(Rickettsia raoultii、Candidatus Rickettsia tarasevichiae、Rickettsia heilongjiangensis),也同时证实了嗜吞噬细胞无形体(Anaplasma phagocytophilum)、巴贝西原虫(Babesia spp.)、巴尔通体(Bartonella spp.)的存在。3、复合感染检测结果共检测到复合感染24例,分别为斑点热群立克次体与莱姆病螺旋体,斑点热群立克次体与嗜吞噬细胞无形体和斑点热群立克次体与巴尔通体,复合感染率为18.4‰(24/1306)。结论:黑龙江口岸媒介蜱类携带多种新发蜱媒病原体,并存在一定程度的复合感染现象,提示黑龙江地区蜱媒疾病的危险程度很高,急需加强黑龙江口岸地区蜱媒传染病及其复合感染的监测与防控技术研究,进而建立科学合理的新发蜱媒疾病预警监测和防控技术体系。
[Abstract]:Objective: To study the distribution, carrying and compound infection of new infectious pathogens in Mishan, Fenhe River, Jiayin, Dongning, Hualin, Tongjiang, Sunke, Lubei, Mudanjiang, Hei River, Raohe 11 port in Heilongjiang area. Method: From April, 2014 to October 2015, the ecological and biological history data of forest vegetation and medium-term disease in Heilongjiang port were integrated. Samples were collected by the cloth flag method. After morphological identification, the samples were numbered according to the area and type, and only one was detected. The method comprises the following steps of: carrying out independent subpackaging, numbering on the pipe wall, placing in a sterile steel ball, grinding the pellets into powder at a low temperature environment by using a grinder, storing at 4 DEG C, extracting tissue DNA according to the operation steps of the kit, establishing five new infectious diseases pathogens (Borrelia burgdorferi, carrying out specific fragment amplification on all samples by using a PCR or a nested PCR method, selecting a PCR positive fragment to carry out sequencing by using a PCR or a nested PCR method, The obtained sequences were compared with the online (NCBI) sequence, and analyzed by tree analysis to determine and analyze the geographic and characteristic distribution of pathogens in the media. Results: A total of 1306 samples were collected by classification and identification. Among the 1306 samples tested, there were only 25.42% of the total population, 25.42% of the whole trench, 23. 58% of the total channel, and 3. 14% of the Japanese blood. The result of the pathogen detection was in 1306 samples tested. A total of 1cases of Leishmania positive samples, 528 spotted fever group rickettsia positive samples, 11 cases of Bartonella positive samples, 31 cases of phagocyte-free positive samples, 11 cases of Lyme disease spiral positive samples were detected, and the infection rates were 0. 8%, 404. 3, 8. 4, 23. 7 and 8. 4%, respectively. A total of 8 pathogens were detected, including Borrelia afzelii, Borrelia garinii of Borrelia afzelii, Borrelia garinii, Babesia spp. of Borrelia garinii, Babesia spp., Racktsia raoulitii, New Taraswichtsia tabaci, Heilkettsia heilongjianguensis, Heilongjiang Province, respectively. Bartonella spp., phagocyte-free anaplasma phagocytophilum. The results of this study show that there are at least two Borrelia garinii, Borrelia afzelii, and three spot heat groups (Rickettsia raoulitii, Riceidi Rickettsia tarachaichensis, Rickettsia heilongjiangensis) in 11 port areas of Heilongjiang Province, and also confirms that the phagocyte-free body (Anapasma phagocytophilum), Babesia spp.), Bartonella spp.) The complex infection rate was 18. 4% (24/ 1306). Conclusion: There is a certain degree of compound infection in the media-borne diseases in Heilongjiang province. It is suggested that the risk of drug-borne diseases in Heilongjiang province is very high. It is urgent to strengthen the research on the monitoring and prevention and control of the infectious diseases and their complex infections in the area of Heilongjiang, and establish a scientific and reasonable system of early warning monitoring and prevention and control of disease-borne diseases.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R181.3

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