西藏阿里地区细粒棘球蚴人体分离株nad1基因多态性分析

doi:10.12140/j.issn.1000-7423.2020.01.003

中国寄生虫学与寄生虫病杂志 ›› 2020, Vol. 38 ›› Issue (1): 17-22.doi: 10.12140/j.issn.1000-7423.2020.01.003

西藏阿里地区细粒棘球蚴人体分离株nad1基因多态性分析

魏玉环¹, 刘华¹, 李武军², 赵海³, 胡媛^1,*, 曹建平¹

1 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;
2 西安医学院第一附属医院普通外科,西安 710077;
3 陕西省人民医院检验科,西安 710068

收稿日期:2019-07-21 出版日期:2020-02-28 发布日期:2020-03-19
通讯作者: 胡媛,E-mail：huyuan@nipd.chinacdc.cn
作者简介:魏玉环(1993-) 女,硕士研究生,从事寄生虫病防控研究。E-mail：weiyh688@126.com
基金资助:
国家科技重大专项(No. 2018ZX10102001-002-004); 国家自然科学基金(No. 81772225,No. 81971969); 西藏阿里地区自然科学基金（No. akkczrjj20180202）

Analysis of nad1 gene polymorphisms of Echinococcus granulosus isolates from humans in Ali region of Tibet

WEI Yu-huan¹, LIU Hua¹, LI Wu-jun², ZHAO Hai³, HU Yuan^1,*, CAO Jian-ping¹

1 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Chinese Center for Tropical Diseases Research; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology; Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China;
2 General Surgery, First Affiliated Hospital of Xi’an Medical College, Xi’an 710077, China;;
3 Laboratory Department of Shaanxi People’s Hospital, Xi’an 710068, China;

Received:2019-07-21 Online:2020-02-28 Published:2020-03-19
Contact: E-mail：huyuan@nipd.chinacdc.cn
Supported by:
Supported by National Science and Technology Major Project (No. 2018ZX10102001-002-004), National Natural Science Foundation of China (No. 81772225,No. 81971969), and Natural Science Foundation of Ali Region, Tibet (No. akkczrjj20180202)

摘要/Abstract

摘要： 目的了解西藏阿里地区细粒棘球蚴人体分离株的优势基因型及遗传变异情况,为细粒棘球绦虫的溯源及阿里地区棘球蚴病预防控制策略的制定提供支持。方法收集阿里地区某医院2017年棘球蚴病患者病灶切除样品,提取DNA,PCR扩增线粒体NADH脱氢酶1（nad1）基因,扩增产物测序后用BLAST、ClustalX 1.83和MEGA 7.0软件进行同源性和系统进化分析。下载全国细粒棘球蚴人体分离株的nad1基因,利用DnaSP6分析单倍型;利用NetWork软件制作中国细粒棘球绦虫nad1基因的单倍型网络图。结果共收集80例细粒棘球蚴病患者病灶切除样品,其中38份样品PCR扩增出约550 bp的特异性条带。测序分析结果显示,4份样品为细粒棘球绦虫G6基因型,与蒙古人来源的序列（MH300971.1）一致性为99.8%;其余34份样品均为细粒棘球绦虫G1基因型,与标准序列（AF297617.1）相比,其535位的C碱基突变为G碱基,与阿尔及利亚人来源序列（MG672293.1）的一致性为100%。MEGA 7.0软件分析38份样品的序列,结果显示,T、C、A和G碱基占比依次为44.3%~46.5%、7.7%~9.1%、19.8%~21.5%和25.7%~26.0%。我国已报道的细粒棘球绦虫nad1基因共有9个单倍型,单倍型多样性为0.47,核酸多样性为0.19。单倍型网络图显示,H4为我国细粒棘球绦虫nad1基因主要的单倍型。结论细粒棘球绦虫G1、G6基因型是西藏阿里地区的主要基因型,资料分析表明H4为我国流行的细粒棘球绦虫nad1基因的主要单倍型。

关键词: 细粒棘球蚴, 基因多态性, nad1基因, 单倍型

Abstract: Objective To understand the dominant genotypes and genetic variations of Echinococcus granulosus isolates from humans in Ali region, in order to provide support for tracing the origin of E. granulosus and formulating strategies for prevention and control of echinococcosis in Ali region. Methods Specimens resected from hydatidosis patients in 2017 in a hospital in Ali area were used for DNA extraction. Mitochondrial NADH dehydrogenase 1 (nad1) gene was amplified by PCR. The PCR products were sequenced and analyzed by BLAST, ClustalX 1.83 and MEGA 7.0 softwares for homology and phylogenetic analyses. The nad1 gene sequences of E. granulosus in China was downloaded and the haplotypes were analyzed by DnaSP6. The haplotype network map of nad1 gene of E. granulosus in China was drawn by the NetWork software. Results A total of 80 specimens of hydatidosis were collected, and 38 resulted in a specific band around 550 bp after PCR amplification. Sequencing analysis revealed that E. granulosus in 4 cases had the G6 genotype, which was 99.81% homogeneous with the sequence of Mongolian origin (MH300971.1). The remaining 34 cases revealed G1 genotype of E. granulosus, which had a C-to-G mutation at position 535 as compared to the standard sequence (AF297617.1) and was 100% homogeneous with that of the Algerian origin (MG672293.1). Analysis of the 38 samples with MEGA 7.0 software showed that the bases T, C, A and G constituted 44.3%-46.5%, 7.7%-9.1%, 19.8%-21.5% and 25.7%-26.0%, respectively. The reported E. granulosus nad1 gene in China had 9 haplotypes, with a haplotype diversity of 0.47 and nucleotide diversity of 0.19. The haplotype network map showed that H4 was the main haplotype for nad1 gene in China. Conclusion G1 and G6 are the genotypes of E. granulosus nad1 gene in Ali region. The data analysis indicates that H4 is the major haplotype for E. granulosus nad1 gene in China.

Key words: Echinococcus granulosus, Genetic polymorphism, nad1 gene, Haplotype

中图分类号:

R383.33

魏玉环, 刘华, 李武军, 赵海, 胡媛, 曹建平. 西藏阿里地区细粒棘球蚴人体分离株nad1基因多态性分析[J]. 中国寄生虫学与寄生虫病杂志, 2020, 38(1): 17-22.

WEI Yu-huan, LIU Hua, LI Wu-jun, ZHAO Hai, HU Yuan, CAO Jian-ping. Analysis of nad1 gene polymorphisms of Echinococcus granulosus isolates from humans in Ali region of Tibet[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2020, 38(1): 17-22.

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西藏阿里地区细粒棘球蚴人体分离株nad1基因多态性分析

Analysis of nad1 gene polymorphisms of Echinococcus granulosus isolates from humans in Ali region of Tibet

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