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我国不同利什曼病流行区利什曼原虫分离株核糖体内转录间隔区1的多态性分析
其他题名Polymorphism analysis of internal transcribed spacer 1 of Leishmania isolates from different endemic areas in China
程芳洲; 高春花; 杨玥涛; 汪俊云
来源期刊中国寄生虫学与寄生虫病杂志
ISSN1000-7423
出版年2019
卷号37期号:2页码:196-201
中文摘要目的分析我国不同利什曼病流行区的利什曼原虫分离株核糖体内转录间隔区1(ITS-1)序列的多态性。方法实验室培养10个分离自我国不同利什曼病流行区的利什曼原虫分离株,包括四川/甘肃山丘型内脏利什曼病流行区的SC6和Cy 分离株,新疆喀什人源型内脏利什曼病流行区的801和KS-2分离株,新疆伽师荒漠型内脏利什曼病流行区的JIASHI-1、JIASHI-5、XJ771分离株以及新疆克拉玛依皮肤利什曼病流行区的KXG-Xu、 KXG-Liu和KXG-927分离株。收集利什曼原虫前鞭毛体,提取DNA,PCR 扩增各利什曼原虫分离株的ITS-1并送测序,用GENEDOC 软件对ITS-1序列进行比对分析,从GenBank 下载利什曼原虫其他分离株的ITS-1序列,构建系统进化树,分析我国利什曼原虫分离株的亲缘关系。结果我国10个利什曼原虫分离株均扩增出单一的条带。序列分析结果显示,801和KS-2分离株的ITS-1片段为316bp,与MHOM/IN/80/DD8参照株比较其碱基突变率为0;JIASHI-1、JIASHI-5和XJ771分离株为313bp,碱基突变率为1.6%;KXG-Xu、KXG-Liu和KXG-927分离株为312bp,碱基突变率为1.6%;Cy和SC6分离株为318bp,碱基突变率为0.6%。系统进化树显示,这10个利什曼原虫分离株ITS-1序列聚集成2个群和3个亚群,其中JIASHI-1、JIASHI-5和XJ771分离株聚集为A 亚群,KXG-Xu、KXG-Liu和KXG-927分离株聚集为B 亚群,A和B 亚群聚集于群Ⅰ,为婴儿利什曼原虫;Cy、 SC6、801和KS2分离株聚集为C 亚群,属群Ⅱ,为杜氏利什曼原虫。结论我国不同利什曼病流行区的利什曼原虫分离株的ITS-1序列存在多态性。
英文摘要Objective The polymorphism of the internal transcribed spacer 1 (ITS-1) sequence of Leishmania isolates from different endemic areas in China was analyzed to understand the genetic variation and relationship among different Leishmania isolates.Methods Total 10 Chinese Leishmania isolates,including SC6 and Cy isolates from endemic areas of mountainous visceral leishmaniasis in Sichuan/Gansu Province (Leishmania donovani);801 and KS-2 isolates from endemic areas of anthroponotic visceral leishmaniasis in Kashi,Xinjiang (L.donovani);JIASHI-1,JIASHI-5 and XJ771 isolates from endemic areas of desert-type visceral leishmaniasis in Jiashi,Xinjiang (L.infantum);KXG-Xu,KXG-Liu and KXG-927 isolates from endemic areas of cutaneous leishmaniasis in Karamay,Xinjiang(L.infantum),were maintained in the laboratory culture.The genomic DNAs were extracted from the cultured promastigotes and the ITS-1 was PCR amplified using gene-specific primers.The PCR products of amplified ITS-1 were sequenced and the obtained sequences from different isolates were aligned with reference ITS-1 sequences of Leishmania deposited in GenBank using GENEDOC software.The phylogenetic tree was constructed based on their genetic variation and relationship.Results A single band of ITS-1 with size of about 310 bp was amplified from all 10 Chinese Leishmania isolates.The sequence analysis identified that ITS-1 amplified from anthroponotic L.donovani 801 and KS-2 isolates contained 316 bp with sequence identical to ITS-1 sequence from L.donovani reference strain MHOM/IN/80/DD8 without any mutation.The ITS-1 amplified from desert-type L.infantum JIASHI-1,JIASHI-5 and XJ771 isolates were 313 bp with 1.6% sequence variation.The ITS-1 amplified from cutaneous L.infantum KXGXu,KXG-Liu and KXG-927 isolates were 312 bp with 1.6% sequence variation.The ITS-1 amplified from mountainous L.donovani Cy and SC6 isolates were 318 bp with 0.6% sequence variation.Phylogenetic analysis of the ITS-1 sequences showed that the 10 Leishmania isolates were branched into three subgroups and two clusters according to the different species and endemic areas.The L.infantum isolates from Jiashi,Xinjiang (JIASHI-1,JIASHI- 5 and XJ771) were closely related and clustered as subgroup A.The L.infantum isolates from Karamay,Xinjiang(KXG-Xu,KXG-Liu and KXG-927) are clustered as subgroup B.Subgroup A and B are closely related to L.infantum reference as cluster Ⅰ.The L.donovani isolates Cy,SC6,801 and KS2 are closely related as subgroup C,fallen into cluster Ⅱof L.donovani.Conclusion ITS-1 sequences of Leishmania isolates from different leishma niasis endemic areas in China showed different polymorphism related to different Leishmania species and endemic areas.
中文关键词利什曼病 ; 利什曼原虫 ; 分离株 ; 内转录间隔区1 ; 基因分型 ; 中国
英文关键词Leishmaniasis Leishmania Isolate Internal transcribed spacer 1 Genotyping China
语种中文
国家中国
收录类别CSCD
WOS类目MEDICINE GENERAL INTERNAL
WOS研究方向General & Internal Medicine
CSCD记录号CSCD:6495698
资源类型期刊论文
条目标识符http://119.78.100.177/qdio/handle/2XILL650/239740
作者单位中国疾病预防控制中心寄生虫病预防控制所, 国家热带病研究中心;;世界卫生组织热带病合作中心;;科技部国家级热带病国际联合研究中心;;卫生部寄生虫病原与媒介生物学重点实验室, 上海 200025, 中国
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程芳洲,高春花,杨玥涛,等. 我国不同利什曼病流行区利什曼原虫分离株核糖体内转录间隔区1的多态性分析[J],2019,37(2):196-201.
APA 程芳洲,高春花,杨玥涛,&汪俊云.(2019).我国不同利什曼病流行区利什曼原虫分离株核糖体内转录间隔区1的多态性分析.中国寄生虫学与寄生虫病杂志,37(2),196-201.
MLA 程芳洲,et al."我国不同利什曼病流行区利什曼原虫分离株核糖体内转录间隔区1的多态性分析".中国寄生虫学与寄生虫病杂志 37.2(2019):196-201.
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