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科威特呼吸道感染患者呼吸道样本中病毒多样性的宏基因组分析。

Metagenomic analysis of viral diversity in respiratory samples from patients with respiratory tract infections in Kuwait.

机构信息

Virology Unit, Department of Microbiology, Faculty of Medicine, Kuwait University, Safat, Kuwait.

Research Core Facility and OMICS Research Unit, Faculty of Medicine, Kuwait University, Safat, Kuwait.

出版信息

J Med Virol. 2018 Mar;90(3):412-420. doi: 10.1002/jmv.24984. Epub 2017 Nov 11.

Abstract

A metagenomic approach based on target independent next-generation sequencing has become a known method for the detection of both known and novel viruses in clinical samples. This study aimed to use the metagenomic sequencing approach to characterize the viral diversity in respiratory samples from patients with respiratory tract infections. We have investigated 86 respiratory samples received from various hospitals in Kuwait between 2015 and 2016 for the diagnosis of respiratory tract infections. A metagenomic approach using the next-generation sequencer to characterize viruses was used. According to the metagenomic analysis, an average of 145, 019 reads were identified, and 2% of these reads were of viral origin. Also, metagenomic analysis of the viral sequences revealed many known respiratory viruses, which were detected in 30.2% of the clinical samples. Also, sequences of non-respiratory viruses were detected in 14% of the clinical samples, while sequences of non-human viruses were detected in 55.8% of the clinical samples. The average genome coverage of the viruses was 12% with the highest genome coverage of 99.2% for respiratory syncytial virus, and the lowest was 1% for torque teno midi virus 2. Our results showed 47.7% agreement between multiplex Real-Time PCR and metagenomics sequencing in the detection of respiratory viruses in the clinical samples. Though there are some difficulties in using this method to clinical samples such as specimen quality, these observations are indicative of the promising utility of the metagenomic sequencing approach for the identification of respiratory viruses in patients with respiratory tract infections.

摘要

基于靶标独立的下一代测序的宏基因组方法已成为在临床样本中检测已知和新型病毒的已知方法。本研究旨在使用宏基因组测序方法来描述呼吸道感染患者的呼吸道样本中的病毒多样性。我们研究了 2015 年至 2016 年间科威特各医院送检的 86 例呼吸道样本,用于呼吸道感染的诊断。使用下一代测序仪对宏基因组方法进行了病毒特征分析。根据宏基因组分析,平均鉴定出 145019 个读数,其中 2%的读数来源于病毒。此外,病毒序列的宏基因组分析揭示了许多已知的呼吸道病毒,在 30.2%的临床样本中检测到这些病毒。在 14%的临床样本中检测到非呼吸道病毒的序列,而在 55.8%的临床样本中检测到非人类病毒的序列。病毒的平均基因组覆盖率为 12%,其中呼吸道合胞病毒的最高基因组覆盖率为 99.2%,而 torque teno midi 病毒 2 的最低覆盖率为 1%。我们的结果表明,在检测临床样本中的呼吸道病毒时,多重实时 PCR 和宏基因组测序之间有 47.7%的一致性。尽管该方法在临床样本中存在一些困难,例如标本质量,但这些观察结果表明宏基因组测序方法在鉴定呼吸道感染患者的呼吸道病毒方面具有广阔的应用前景。

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