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从猪带绦虫全基因组分析中鉴定和表征微卫星标记

Identification and Characterization of Microsatellite Markers Derived from the Whole Genome Analysis of Taenia solium.

作者信息

Pajuelo Mónica J, Eguiluz María, Dahlstrom Eric, Requena David, Guzmán Frank, Ramirez Manuel, Sheen Patricia, Frace Michael, Sammons Scott, Cama Vitaliano, Anzick Sarah, Bruno Dan, Mahanty Siddhartha, Wilkins Patricia, Nash Theodore, Gonzalez Armando, García Héctor H, Gilman Robert H, Porcella Steve, Zimic Mirko

机构信息

Laboratorio de Bioinformatica y Biologia Molecular, Laboratorios de Investigación y Desarrollo, Facultad de Ciencias y Filosofía, Universidad Peruana Cayetano Heredia, Lima, Peru.

Genomics Unit, Research Technologies Section, Rocky Mountain Laboratories, NIAID, NIH, Hamilton, Montana, United States of America.

出版信息

PLoS Negl Trop Dis. 2015 Dec 23;9(12):e0004316. doi: 10.1371/journal.pntd.0004316. eCollection 2015 Dec.

Abstract

BACKGROUND

Infections with Taenia solium are the most common cause of adult acquired seizures worldwide, and are the leading cause of epilepsy in developing countries. A better understanding of the genetic diversity of T. solium will improve parasite diagnostics and transmission pathways in endemic areas thereby facilitating the design of future control measures and interventions. Microsatellite markers are useful genome features, which enable strain typing and identification in complex pathogen genomes. Here we describe microsatellite identification and characterization in T. solium, providing information that will assist in global efforts to control this important pathogen.

METHODS

For genome sequencing, T. solium cysts and proglottids were collected from Huancayo and Puno in Peru, respectively. Using next generation sequencing (NGS) and de novo assembly, we assembled two draft genomes and one hybrid genome. Microsatellite sequences were identified and 36 of them were selected for further analysis. Twenty T. solium isolates were collected from Tumbes in the northern region, and twenty from Puno in the southern region of Peru. The size-polymorphism of the selected microsatellites was determined with multi-capillary electrophoresis. We analyzed the association between microsatellite polymorphism and the geographic origin of the samples.

RESULTS

The predicted size of the hybrid (proglottid genome combined with cyst genome) T. solium genome was 111 MB with a GC content of 42.54%. A total of 7,979 contigs (>1,000 nt) were obtained. We identified 9,129 microsatellites in the Puno-proglottid genome and 9,936 in the Huancayo-cyst genome, with 5 or more repeats, ranging from mono- to hexa-nucleotide. Seven microsatellites were polymorphic and 29 were monomorphic within the analyzed isolates. T. solium tapeworms were classified into two genetic groups that correlated with the North/South geographic origin of the parasites.

CONCLUSIONS/SIGNIFICANCE: The availability of draft genomes for T. solium represents a significant step towards the understanding the biology of the parasite. We report here a set of T. solium polymorphic microsatellite markers that appear promising for genetic epidemiology studies.

摘要

背景

猪带绦虫感染是全球成人后天性癫痫最常见的病因,也是发展中国家癫痫的主要病因。更好地了解猪带绦虫的遗传多样性将改善流行地区的寄生虫诊断和传播途径,从而有助于设计未来的控制措施和干预手段。微卫星标记是有用的基因组特征,能够在复杂病原体基因组中进行菌株分型和鉴定。在此,我们描述了猪带绦虫中微卫星的鉴定和特征,提供的信息将有助于全球控制这种重要病原体的努力。

方法

为进行基因组测序,分别从秘鲁的万卡约和普诺收集猪带绦虫囊肿和节片。使用下一代测序(NGS)和从头组装,我们组装了两个草图基因组和一个混合基因组。鉴定了微卫星序列,并选择其中36个进行进一步分析。从秘鲁北部的通贝斯收集了20个猪带绦虫分离株,从南部的普诺收集了20个。通过多毛细管电泳确定所选微卫星的大小多态性。我们分析了微卫星多态性与样本地理来源之间的关联。

结果

猪带绦虫混合基因组(节片基因组与囊肿基因组组合)的预测大小为111 MB,GC含量为42.54%。共获得7979个重叠群(>1000 nt)。我们在普诺节片基因组中鉴定出9129个微卫星,在万卡约囊肿基因组中鉴定出9936个,重复次数为5次或更多,范围从单核苷酸到六核苷酸。在所分析的分离株中,7个微卫星是多态性的,29个是单态性的。猪带绦虫绦虫被分为两个遗传组,这与寄生虫的南北地理来源相关。

结论/意义:猪带绦虫草图基因组的可用性是朝着了解该寄生虫生物学迈出的重要一步。我们在此报告了一组猪带绦虫多态性微卫星标记,这些标记在遗传流行病学研究中似乎很有前景。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11eb/4689449/03bbdf55c4da/pntd.0004316.g001.jpg

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