吉林延边地区蜱复合感染新塔拉塞维奇立克次体及SFTSV调查研究

李基旭; 赵鑫; 金光星; 金光俊; 马桂君; 金京男; 全圣焕; Shin-HyeongCho

doi:10.16462/j.cnki.zhjbkz.2019.05.014

吉林延边地区蜱复合感染新塔拉塞维奇立克次体及SFTSV调查研究

doi: 10.16462/j.cnki.zhjbkz.2019.05.014

1.
133001 延吉, 吉林省延边州疾病预防控制中心病媒科
2.
133200 汪清, 吉林省汪清县疾病预防控制中心传染病控制科
3.
133600 安图, 吉林省安图县疾病预防控制中心传染病控制科
4.
133300 珲春, 吉林省珲春市疾病预防控制中心监测科
5.
133700 敦化, 吉林省敦化市疾病预防控制中心传染病控制科
6.
133001 延吉, 吉林省延吉市疾病预防控制中心地方病科
7.
133500 和龙, 吉林省和龙市疾病预防控制中心传染病控制科
8.
363951 清州, 韩国疾病预防控制中心国立保健研究院媒介分析科

基金项目: 韩国疾病预防控制中心国立保健研究院国际感染性疾病检测体系援助项目

详细信息

通讯作者:
Shin-Hyeong Cho, E-mail: cho4u@korea.kr

中图分类号: R181;R51
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出版历程
- 收稿日期: 2018-02-23
- 修回日期: 2018-08-09
- 刊出日期: 2019-05-10

Investigation of free ticks carrying CRT and compound infection with SFTSV in Yanbian area of Jilin Province

1.
Center for Disease Control and Prevention Division of Vector Control, Yanbian 133001, China
2.
Center for Disease Control and Prevention Division of Infectious Disease Control, WangQing 133200, China
3.
Center for Disease Control and Prevention Division of Infectious Disease Dontrol, AnTu 133200, China
4.
Center for Disease Control and Prevention Division of Health Monitoring, HunChun 133300, China
5.
Center for Disease Control and Prevention Division of Infectious Disease Control, DunHua 133700, China
6.
Center for Disease Control and Prevention Division of Local Disease Surveillance, YanJi 133000, China
7.
Center for Disease Control and Prevention Division of Infectious Disease Control, HeLong, Yanbian 133500, China
8.
National Research Institute of Health, South of Korea Division of vectors and parasitic Diseases, Cheongju 363951, Korea

Funds: International Laboratory Installation and Collaborative Research on Neglected Tropical and Vector-borne Diseases, KCDC

More Information

Corresponding author: Shin-Hyeong Cho, E-mail: cho4u@korea.kr

摘要

摘要: 目的为了解吉林省延边地区蜱类新塔拉塞维奇立克次体(candidatus rickettsia tarasevichiae，CRT)和复合感染新布尼亚病毒(sever fever thrombocytopenia syndrome virus，SFTSV)的感染情况。方法采集吉林省延边朝鲜族自治州(延边州)延吉、汪清、珲春、敦化、安图、和龙6个县游离蜱，进行形态学分类，通过巢式多聚核苷酸链式反应(nested polymerase chain reaction，nested PCR)和实时荧光定量多聚核苷酸链式反应(real-time quantitative polymerase chain reaction，RT-qPCR)方法检测蜱体内CRT和SFTSV。对其CRT进行目标DNA测序，分析同源性和系统进化关系。结果此研究共采集各种蜱类1 032只，分别为全沟硬蜱(ixodes persulcatus)(35.56%)、森林革蜱(dermacentor silvarum)(20.64%)、日本血蜱(haemaphysalis japonica)(20.45%)、长角血蜱(haemaphysalis longicornis)(10.47%)、嗜群血蜱(haemaphysalis concinna)(8.33%)以及其他种类(4.55%)。全沟硬蜱、长角血蜱、日本血蜱、森林革蜱检测出CRT，最低感染率(minimum infection rate per 100 ticks，MIR)为10.47%。嗜群血蜱、日本血蜱、全沟硬蜱、长角硬蜱、森林革蜱检测出SFTSV，最低感染率为2.52%。全沟硬蜱、日本血蜱、森林革蜱3种蜱存在CRT和SFTSV复合感染现象，复合感染MIR为1.26%。本研究CRT ompA和17kDa基因序列与中国河南信阳株(KX365196.1)序列同源性达100%。系统进化分析，延边地区蜱CRT ompA基因序列和该河南信阳株基因序列成为一簇，而17kDa基因则形成独立分支。结论本研究首次从吉林省延边地区游离蜱中检测出CRT，全沟硬蜱为主要媒介宿主。因此，可以明确该地区为新塔拉塞维奇立克次体病自然疫源地。同时发现，延边地区蜱类存在CRT与SFTSV复合感染的现象。因此，在该地区必须加强新塔拉塞维奇立克次体病和发热伴血小板减少综合征(sever fever thrombocytopenia syndrome，SFTS)疫情监测。
- 蜱 /
- 新塔拉塞维奇立克次体 /
- 新布尼亚病毒 /
- 复合感染
Abstract: Objective To understand the condition of tick carrying CRT(Candidatus rickettsia tarasevichiae) and compound infection with SFTSV(Sever fever thrombocytopenia syndrome virus) in Yanbian area of Jilin province. Methods Free ticks were collected from 6 counties including Yanji, Wangqing, Hunchun, Dunhua, Antu and Helong in the Korean Autonomous Prefecture of Jilin Province(Yanbian Prefecture) by using the flagging, and they were classified morphologically. The CRT and SFTSV in ticks were detected by Nest PCR(Nested polymerase chain reaction) and Real Time RT-PCR(Real-time Quantitative polymerase chain reaction) methods. Target DNA sequencing was performed for CRT positive products, and the homology of gene sequence and phylogenetic relationship were analyzed. Results A total of 1032 ticks were collected in this study, including ixodes persulcatus (35.56%) and dermacenter silvarum (20.64%). haemaphysalis japonica (20.45%), haemaphysalis longicornis (10.47%), haemaphysalis concinna (8.33%), others(4.55%). The CRT was detected from the Ixodes persulcatus, Haemaphysalis longicornis, Haemaphysalis japonica Dermacentor silvarum. The MIR(Minimum infection rate per 100 ticks) of CRT was 10.47%.The SFTSV was detected from the Haemaphysalis-concinna, Haemaphysalis-japonica, Ixodes-persulcatus, Haemaphysalis-longicornis, Dermacentor-silvarum. The MIR of SFTSV was 2.52%. Three species of ticks, including Ixodes persulcatus(2.45%), Haemaphysalis japonica(1.42%), and dermacentor silvarum(0.47%), had CRT and SFTSV compound infections, and the MIR of two pathogens compound infections was 1.26%. In this study, the gene sequence of CRT positive PCR products ompA and 17kDa with nucleotide sequence of Xinyang plant of HeNan XinYang strain (KX365196.1), had homology of 100%. Phylogenetic analysis showed that CRT ompA and the HeNan XinYang strain (KX365196.1) gene sequences formed a cluster in Yanbian, while the 17 kDa gene formed an independent branch. Conclusions For the first time, CRT was detected from free ticks in Yanbian area of Jilin Province, and it was found that Ixodes persulcatus may be the main medium of transmission of the pathogen. At the same time, it was found that CRT and SFTSV have compound infection in ticks of Yanbian area. Therefore, it can be clearly identified that Yanbian area in Jilin Province is the natural source of CRT, and there are two pathogenic compound infections in the local ticks.
- Tick /
- Candidatus Rickettsia tarasevichiae /
- Sever fever thrombocytopenia syndrome virus /
- Compound infection

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图 1 CRT ompA基因扩增结果

M: DNA标志物；3, 4:阳性样品(346bp)；1, 2:阴性样品；NC: 阴性对照

Figure 1. The amplification of CRT ompA by nested PCR

下载: 全尺寸图片幻灯片

图 2 CRT 17kDa基因扩增结果

M: DNA标志物；S1-S5:阳性样品(395bp)；NC: 阴性对照. by nested PCR

Figure 2. The amplification of CRT 17kDa

下载: 全尺寸图片幻灯片

图 3 延边地区游离蜱CRT和SFTSV检测情况

C: 检测出CRT阳性; S: 检测出SFTSV阳性; C & S: 检测出CRT和SFTSV阳性; CS: 检测出蜱复合感染CRT和SFTSV

Figure 3. The detection of CRT and SFTSV in free tick in YanBian area

下载: 全尺寸图片幻灯片

图 4 基于ompA基因片段的遗传进化分析

注：近邻相接法(neighbor-joining method); Bootstrap检验，重复检验1 000次。

Figure 4. Phylogenetic analysis of ompA gene of CRT

Method: neighbor-joining; Test: Bootstrap; Number of Test : 1 000。

下载: 全尺寸图片幻灯片

图 5 基于CRT 17kDa基因片段的遗传进化分析

注：近邻相接法(neighbor-joining method); Bootstrap检验，重复检验1 000次。Method: neighbor-joining; Test: Bootstrap; Number of Test : 1 000。

Figure 5. Phylogenetic analysis of 17kDa gene of CRT

下载: 全尺寸图片幻灯片

表 1 扩增CRT ompA基因引物序列

Table 1. The List of primer for CRT ompA

引物名称	序列(5'-3')	目标片段大小
Rr190.70p	ATGGCGAATATTTCTCCAAAA	346bp
Rr190.602n	AGTGCAGCATTCGC TCCCCCT
190.70-38s1	AAAACCG CTTTATTCACC
190.602-384r1	GGCAAC AAGTTACCTCCT

下载: 导出CSV

表 2 扩增CRT 17kDa基因引物序列

Table 2. The List of primer for CRT 17kDa

引物名称	序列(5'-3')	目标片段大小
17K3	GCTTTACAA AATTCTAAAAACCATATA	395 bp
17K5	TGTCTATCAATTCACAACTTGCC
17KD113s1	ATTGTCCGTCAGGTTGGC
17KD408r1	CGGGCGGTATGAATAAGC

下载: 导出CSV

表 3 延边地区蜱CRT和SFTSV检测结果[n(%)]

Table 3. The detection of CRT and SFTSV in free tick in YanBian area[n(%)]

种类	数量(n)	CRT	SFTSV
种类	数量(n)	最低感染率(MIR)	最低感染(MIR)
森林革蜱	213	11(5.16)	1(0.47)
日本血蜱	211	12(5.69)	8(3.79)
全沟硬蜱	367	70(19.07)	11(3.00)
长角血蜱	108	13(12.04)	1(0.93)
嗜群血蜱	86	0(0.00)	5(5.81)
其他	47	2(4.26)	0(0.00)
合计	1032	108(10.47)	26(2.52)

下载: 导出CSV

表 4 延边地区蜱CRT和SFTSV复合感染检测结果[n(%)]

Table 4. The detection of Co-infection of CRT and SFTSV in ticks in YanBian area[n(%)]

种类	数量(个)	最低感染率(MIR)
全沟硬蜱	367	9(2.45)
日本血蜱	211	3(1.42)
森林革蜱	213	1(0.47)
长角血蜱	108	0(0.00)
嗜群血蜱	86	0(0.00)
其他	47	0(0.00)
合计	1032	13(1.26)

下载: 导出CSV

表 5 不同蜱种CRT阳性率比较[n(%)]

Table 5. The comparison of CRT positive rates among different ticks[n(%)]

蜱种	采样组数(个)	阳性率
全沟硬蜱	80	70(87.50)
长角血蜱	16	13(81.25)
日本血蜱	46	12(26.09)
森林革蜱	44	11(25.00)
嗜群血蜱	18	0(0.00)

下载: 导出CSV

表 6 蜱CRT基因GenBank提交序列

Table 6. The accession number of CRT gene from ticks in YanBian area

样品名称	采集地	ompA		17kDa		媒介宿主
样品名称	采集地	序列号	长度(bp)	序列号	长度(bp)	媒介宿主
YBWQ88/2016	汪清	MG888727	346	MG844524	380	全沟硬蜱
YBWQ93/2016	汪清	MG888728	346	MG844525	385	全沟硬蜱
YBWQ374/2016	汪清	MG888729	348	MG844526	391	全沟硬蜱

下载: 导出CSV

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