{"title":"利用基于 EmsB 微卫星的新一代测序技术，对动物和人类包虫囊肿组织样本中的感性棘球蚴进行基因分型。","authors":"Suheir Ereqat, Amer Al-Jawabreh, Hanan Al-Jawabreh, Abedelmajeed Nasereddin","doi":"10.14411/fp.2024.014","DOIUrl":null,"url":null,"abstract":"<p><p>Echinococcus granulosus (Batsch, 1786), a cestode of the Teniidae family, causes human cystic echinococcosis (CE) also known as hydatid disease. Echinococcus granulosus sensu lato includes the G1, G3, G4, G5, G6/7 and G8/10 genotypes which are known to cause human CE. This study aimed to differentiate genotypes of E. granulosus s.l. complex by employing EmsB, a tandemly repeated multilocus microsatellite, using next-generation sequencing (MIC-NGS). Human and animal histopathology-confirmed hydatid cyst tissue samples and reference DNA samples of E. granulosus G1, G3, G4, G5, G6/7 and G10 underwent MIC-NGS assay with custom primers amplifying a 151 bp EmsB DNA fragment. NGS data were analysed using online Galaxy analysis pipeline, a phylogenetic tree was constructed by MEGA software, and haplotype networking was performed with PopArt 1.7. All sixty samples (49 from animals and 11 from humans) included were successfully identified and genotyped with a 100 % success rate. The study showed improved discrimination power to distinguish all study samples including closely related E. granulosus s.s. genotypes G1-G3. The maximum likelihood tree reaffirmed the monophyly of E. granulosus s.l. The median-joining haplotype networking revealed 12 distinct haplotypes. In conclusion, MIC-NGS assay was shown to be sensitive, specific and simple to apply to clinical samples offering a powerful discriminatory tool for the genotyping of E. granulosus s.l.</p>","PeriodicalId":55154,"journal":{"name":"Folia Parasitologica","volume":null,"pages":null},"PeriodicalIF":1.5000,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Use of the EmsB microsatellite-based next generation sequencing for genotyping of Echinococcus granulosus sensu lato in hydatid cyst tissue samples from animals and humans.\",\"authors\":\"Suheir Ereqat, Amer Al-Jawabreh, Hanan Al-Jawabreh, Abedelmajeed Nasereddin\",\"doi\":\"10.14411/fp.2024.014\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Echinococcus granulosus (Batsch, 1786), a cestode of the Teniidae family, causes human cystic echinococcosis (CE) also known as hydatid disease. Echinococcus granulosus sensu lato includes the G1, G3, G4, G5, G6/7 and G8/10 genotypes which are known to cause human CE. This study aimed to differentiate genotypes of E. granulosus s.l. complex by employing EmsB, a tandemly repeated multilocus microsatellite, using next-generation sequencing (MIC-NGS). Human and animal histopathology-confirmed hydatid cyst tissue samples and reference DNA samples of E. granulosus G1, G3, G4, G5, G6/7 and G10 underwent MIC-NGS assay with custom primers amplifying a 151 bp EmsB DNA fragment. NGS data were analysed using online Galaxy analysis pipeline, a phylogenetic tree was constructed by MEGA software, and haplotype networking was performed with PopArt 1.7. All sixty samples (49 from animals and 11 from humans) included were successfully identified and genotyped with a 100 % success rate. The study showed improved discrimination power to distinguish all study samples including closely related E. granulosus s.s. genotypes G1-G3. The maximum likelihood tree reaffirmed the monophyly of E. granulosus s.l. The median-joining haplotype networking revealed 12 distinct haplotypes. In conclusion, MIC-NGS assay was shown to be sensitive, specific and simple to apply to clinical samples offering a powerful discriminatory tool for the genotyping of E. granulosus s.l.</p>\",\"PeriodicalId\":55154,\"journal\":{\"name\":\"Folia Parasitologica\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2024-08-05\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Folia Parasitologica\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.14411/fp.2024.014\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"PARASITOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Folia Parasitologica","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.14411/fp.2024.014","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"PARASITOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
粒棘球蚴（Echinococcus granulosus，Batsch，1786 年）是一种天牛科绦虫，可引起人类囊性棘球蚴病（CE），又称包虫病。普通棘球蚴包括 G1、G3、G4、G5、G6/7 和 G8/10 基因型，已知这些基因型可导致人类囊性棘球蚴病。本研究旨在利用下一代测序技术（MIC-NGS），通过串联重复的多焦点微卫星 EmsB 来区分 E. granulosus s.l. 复合物的基因型。经组织病理学确诊的人类和动物包虫囊肿组织样本以及肉毒梭状芽孢杆菌 G1、G3、G4、G5、G6/7 和 G10 的参考 DNA 样本接受了 MIC-NGS 检测，使用定制引物扩增了 151 bp EmsB DNA 片段。使用在线 Galaxy 分析管道对 NGS 数据进行分析，使用 MEGA 软件构建系统发生树，并使用 PopArt 1.7 进行单倍型网络分析。纳入的所有 60 个样本（49 个来自动物，11 个来自人类）都成功地进行了鉴定和基因分型，成功率为 100%。研究显示，该研究提高了对所有研究样本的鉴别能力，包括与之密切相关的格兰氏阳性大肠杆菌基因型 G1-G3。最大似然树再次证实了 E. granulosus s.l.的单系性。总之，MIC-NGS 检测灵敏、特异且简单，适用于临床样本，是对 E. granulosus s.l. 进行基因分型的有力鉴别工具。
Use of the EmsB microsatellite-based next generation sequencing for genotyping of Echinococcus granulosus sensu lato in hydatid cyst tissue samples from animals and humans.
Echinococcus granulosus (Batsch, 1786), a cestode of the Teniidae family, causes human cystic echinococcosis (CE) also known as hydatid disease. Echinococcus granulosus sensu lato includes the G1, G3, G4, G5, G6/7 and G8/10 genotypes which are known to cause human CE. This study aimed to differentiate genotypes of E. granulosus s.l. complex by employing EmsB, a tandemly repeated multilocus microsatellite, using next-generation sequencing (MIC-NGS). Human and animal histopathology-confirmed hydatid cyst tissue samples and reference DNA samples of E. granulosus G1, G3, G4, G5, G6/7 and G10 underwent MIC-NGS assay with custom primers amplifying a 151 bp EmsB DNA fragment. NGS data were analysed using online Galaxy analysis pipeline, a phylogenetic tree was constructed by MEGA software, and haplotype networking was performed with PopArt 1.7. All sixty samples (49 from animals and 11 from humans) included were successfully identified and genotyped with a 100 % success rate. The study showed improved discrimination power to distinguish all study samples including closely related E. granulosus s.s. genotypes G1-G3. The maximum likelihood tree reaffirmed the monophyly of E. granulosus s.l. The median-joining haplotype networking revealed 12 distinct haplotypes. In conclusion, MIC-NGS assay was shown to be sensitive, specific and simple to apply to clinical samples offering a powerful discriminatory tool for the genotyping of E. granulosus s.l.
期刊介绍：
FOLIA PARASITOLOGICA, issued in online versions, is an international journal that covers the whole field of general, systematic, ecological and experimental parasitology. It publishes original research papers, research notes and review articles. Contributions from all branches of animal parasitology, such as morphology, taxonomy, biology, biochemistry, physiology, immunology, molecular biology and evolution of parasites, and host-parasite relationships, are eligible. Novelty and importance in the international (not local or regional) context are required. New geographical records of parasites, records of new hosts, regional parasite and/or host surveys (if they constitute the principal substance of manuscript), local/regional prevalence surveys of diseases, local/regional studies on epidemiology of well known diseases and of parasite impact on human/animal health, case reports, routine clinical studies and testing of established diagnostic or treatment procedures, will not be considered. One species description will also not be considered unless they include more general information, such as new diagnostic characters, host-parasite associations, phylogenetic implications, etc. Manuscripts found suitable on submission will be reviewed by at least two reviewers.
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