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基于下一代测序的分子方法鉴定莱姆病中的蜱传媒介。

A Next-Generation Sequencing-Based Molecular Approach to Characterize a Tick Vector in Lyme Disease.

机构信息

1 Manipal Academy of Higher Education (MAHE) , Manipal, Karnataka, India .

2 Institute of Bioinformatics , International Technology Park, Bangalore, Karnataka, India .

出版信息

OMICS. 2018 Aug;22(8):565-574. doi: 10.1089/omi.2018.0089.

Abstract

Next-generation sequencing approaches have revolutionized genomic medicine and enabled rapid diagnosis for several diseases. These approaches are widely used for pathogen detection in several infectious diseases. Lyme disease is a tick-borne infectious disease, which affects multiple organs. The causative organism is a spirochete, Borrelia burgdorferi, which is transmitted by ticks. Lyme disease can be treated easily if detected early, but its diagnosis is often delayed or is incorrect leading to a chronic debilitating condition. Current confirmatory diagnostic tests for Lyme disease rely on detection of antigens derived from B. burgdorferi, which are prone to both false positives and false negatives. Instead of focusing only on the human host for the diagnosis of Lyme disease, one could also attempt to identify the vector (tick) and the causative organism carried by the tick. Since all ticks do not transmit Lyme disease, it can be informative to accurately identify the tick from the site of bite, which is often observed by the patient and discarded. However, identifying ticks based on morphology alone requires a trained operator and can still be incorrect. Thus, we decided to take a molecular approach by sequencing DNA and RNA from a tick collected from an individual bitten by the tick. Using next-generation sequencing, we confirmed the identity of the tick as a dog tick, Dermacentor variabilis, and did not identify any pathogenic bacterial sequences, including Borrelia species. Despite the limited availability of nucleotide sequences for many types of ticks, our approach correctly identified the tick species. This proof-of-principle study demonstrates the potential of next-generation sequencing in the diagnosis of tick-borne infections, which can also be extended to other zoonotic diseases.

摘要

下一代测序方法彻底改变了基因组医学,并为多种疾病的快速诊断提供了可能。这些方法广泛用于几种传染病的病原体检测。莱姆病是一种蜱传传染病,可影响多个器官。病原体是一种螺旋体,伯氏疏螺旋体,由蜱传播。如果早期发现,莱姆病很容易治疗,但由于诊断经常被延迟或错误,导致其发展为慢性衰弱性疾病。目前莱姆病的确诊性诊断检测依赖于检测源自伯氏疏螺旋体的抗原,但这些抗原容易出现假阳性和假阴性。除了仅关注人类宿主来诊断莱姆病外,还可以尝试识别携带病原体的媒介(蜱)。由于并非所有蜱都传播莱姆病,因此准确识别来自叮咬部位的蜱很有意义,因为这些蜱通常会被患者观察到并丢弃。然而,仅基于形态学识别蜱需要经过训练的操作人员,而且仍然可能不正确。因此,我们决定采用分子方法,从被蜱叮咬的个体采集的蜱中测序 DNA 和 RNA。使用下一代测序,我们确认了采集的蜱是狗蜱,即璃眼蜱,并未鉴定出任何致病性细菌序列,包括伯氏疏螺旋体。尽管许多类型的蜱的核苷酸序列有限,但我们的方法正确地鉴定了蜱的种类。这项初步研究证明了下一代测序在诊断蜱传感染方面的潜力,该方法也可扩展到其他人畜共患病。

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本文引用的文献

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Lyme borreliosis.莱姆病。
Nat Rev Dis Primers. 2016 Dec 15;2:16090. doi: 10.1038/nrdp.2016.90.
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Incident Tick-Borne Infections in a Cohort of North Carolina Outdoor Workers.北卡罗来纳州一组户外工作者中蜱传感染事件
Vector Borne Zoonotic Dis. 2016 May;16(5):302-8. doi: 10.1089/vbz.2015.1887. Epub 2016 Mar 23.

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