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来自美国东德克萨斯地区临床分离株的严重急性呼吸综合征冠状病毒2(SARS-CoV-2)下一代测序(NGS)数据。

SARS-CoV-2 Next Generation Sequencing (NGS) data from clinical isolates from the East Texas Region of the United States.

作者信息

Carpenter Rob E, Tamrakar Vaibhav K, Almas Sadia, Sharma Aditya, Sharma Rahul

机构信息

Advanta Genetics, 10935 CR 159, Tyler, TX 75703, USA.

University of Texas at Tyler, 3900 University Boulevard, Tyler, TX 75799, USA.

出版信息

Data Brief. 2023 Aug;49:109312. doi: 10.1016/j.dib.2023.109312. Epub 2023 Jun 14.

Abstract

The SARS-CoV-2 virus has evolved throughout the pandemic and is likely to continue evolving into new variants. Some of these variants may affect functional properties, including infectivity, interactions with host immunity, and disease severity. And compromised vaccine efficacy is an emerging concern with every new viral variant. Next-generation sequencing (NGS) has emerged as the tool of choice for discovering new variants and understanding the transmission dynamics of SARS-CoV-2. Deciphering the SARS-CoV-2 genome has enabled epidemiological survivance and forecast of altered etiologically. Clinical presentations of the infection are influenced by comorbidities such as age, immune status, diabetes, and the infecting variant. Thus, clinical management and vaccine efficacy may differ for new variants. For example, some monoclonal antibody treatments are variant-specific, and some vaccines are less efficacious against the omicron and delta variants of SARS-CoV-2. Consequently, determining the local outbreaks and monitoring SARS-CoV-2 Variants of Concern (VOC) is one of the primary strategies for the pandemic's containment. Although next-generation sequencing (NGS) is a gold standard for genomic surveillance and variant discovery, the assays are not approved for variant diagnosis for clinical decision-making. Advanta Genetics, Texas, USA, optimized Illumina COVID-seq protocol to reduce cost without compromising accuracy and validated the Illumina COVID-Seq assay as a Laboratory Developed Test (LDT) according to the guidelines prescribed by the College of American Pathologists (CAP) and Clinical Laboratory Improvement Amendments (CLIA). The whole genome of the virus was sequenced in ( = 161) samples from the East Texas region using the Illumina MiniSeq® instrument and analyzed by using Illumina baseSpace (https://basespace.illumina.com) bioinformatics pipeline. Briefly, the library was prepared by using Illumina COVIDSeq research use only (RUO) kit, and the individual libraries were normalized using the DNA concentration measured by Qubit Flex Fluorometer, and the pooled libraries were sequenced on Illumina MiniSeq® Instrument. Illumina baseSpace application was used for sequencing QC, FASTQ generation, genome assembly, and identification of SARS-CoV-2 variants. This whole genome shotgun project ( = 161) has been deposited at GISAID.

摘要

在整个疫情大流行期间,严重急性呼吸综合征冠状病毒2(SARS-CoV-2)病毒不断进化,并且可能会继续演变成新的变种。其中一些变种可能会影响病毒的功能特性,包括传染性、与宿主免疫的相互作用以及疾病严重程度。而随着每一种新的病毒变种出现,疫苗效力受损正成为一个新的问题。下一代测序(NGS)已成为发现新变种和了解SARS-CoV-2传播动态的首选工具。解读SARS-CoV-2基因组有助于进行流行病学监测以及对病因变化进行预测。感染的临床表现会受到年龄、免疫状态、糖尿病等合并症以及感染变种的影响。因此,针对新变种的临床管理和疫苗效力可能会有所不同。例如,一些单克隆抗体治疗具有变种特异性,一些疫苗对SARS-CoV-2的奥密克戎和德尔塔变种效力较低。因此,确定本地疫情爆发情况并监测SARS-CoV-2关注变种(VOC)是控制疫情大流行的主要策略之一。尽管下一代测序(NGS)是基因组监测和变种发现的金标准,但这些检测方法尚未被批准用于临床决策的变种诊断。美国得克萨斯州的Advanta Genetics公司优化了Illumina COVID-seq方案以降低成本且不影响准确性,并根据美国病理学家协会(CAP)和《临床实验室改进修正案》(CLIA)规定的指南,将Illumina COVID-Seq检测验证为实验室自建检测方法(LDT)。使用Illumina MiniSeq®仪器对来自东得克萨斯地区的161份样本中的病毒全基因组进行测序,并通过Illumina baseSpace(https://basespace.illumina.com)生物信息学管道进行分析。简要来说,使用Illumina COVIDSeq仅供研究使用(RUO)试剂盒制备文库,使用Qubit Flex荧光计测量的DNA浓度对各个文库进行标准化,然后将混合文库在Illumina MiniSeq®仪器上进行测序。使用Illumina baseSpace应用程序进行测序质量控制、FASTQ生成、基因组组装以及SARS-CoV-2变种鉴定。这个全基因组鸟枪法测序项目(n = 161)已存入全球共享流感数据倡议组织(GISAID)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2432/10331279/8ae33633c3a6/gr1.jpg

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