Utility of nanopore sequencing for detecting pathogens in bronchoalveolar lavage fluid from pediatric patients with respiratory failure

IF 1.6 Q4 INFECTIOUS DISEASES Journal of clinical virology plus Pub Date : 2023-06-01 DOI:10.1016/j.jcvp.2023.100154
Makoto Yamaguchi , Kazuhiro Horiba , Kazunori Haruta , Suguru Takeuchi , Takako Suzuki , Yuka Torii , Shinji Kawabe , Sho Wada , Takanari Ikeyama , Yoshinori Ito , Tomoo Ogi , Jun-ichi Kawada
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Abstract

RNA viruses are the most frequent pathogens responsible for respiratory infections, particularly in pediatric patients. Next-generation sequencing, represented by Illumina sequencing, is one of the most comprehensive methods for identifying pathogens. Nanopore sequencing has been used to identify and analyze pathogens with a shorter sequencing time. In this study, we evaluated the utility of nanopore sequencing for the detection of RNA viruses in bronchoalveolar lavage fluid (BALF) of pediatric patients with respiratory failure. Using the seven BALF samples, we first compared the nanopore and Illumina sequencing results. The nanopore sequencing detected the same RNA viruses as the Illumina sequencing. Subsequently, BALF samples from 24 additional pediatric patients with respiratory failure were analyzed by nanopore sequencing, and RNA viral pathogens were detected in 10 out of 24 patients. Among these 10 patients, nanopore sequencing identified the same viral pathogens as detected by the PCR and viral antigen tests in five patients. Furthermore, additional RNA viral pathogens were detected by nanopore sequencing with high genome coverage in five patients that were not detected by PCR and viral antigen tests. In conclusion, nanopore sequencing could comprehensively detect RNA viral pathogens in BALF samples with equivalent sensitivity and genome coverage as Illumina sequencing. This rapid sequencing platform may be more beneficial for detecting RNA viruses in clinical settings.

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应用纳米孔测序检测小儿呼吸衰竭患者支气管肺泡灌洗液中的病原体
RNA病毒是导致呼吸道感染的最常见病原体,尤其是在儿科患者中。以Illumina测序为代表的下一代测序是鉴定病原体最全面的方法之一。纳米孔测序已被用于识别和分析病原体,测序时间更短。在这项研究中,我们评估了纳米孔测序在检测呼吸衰竭儿童患者支气管肺泡灌洗液(BALF)中RNA病毒方面的实用性。使用七个BALF样本,我们首先比较了纳米孔和Illumina测序结果。纳米孔测序检测到与Illumina测序相同的RNA病毒。随后,通过纳米孔测序分析了另外24名呼吸衰竭儿童患者的BALF样本,并在24名患者中的10名中检测到RNA病毒病原体。在这10名患者中,纳米孔测序确定了与5名患者的PCR和病毒抗原检测相同的病毒病原体。此外,通过纳米孔测序在五名患者中检测到了额外的RNA病毒病原体,这些病原体的基因组覆盖率很高,而PCR和病毒抗原测试没有检测到。总之,纳米孔测序可以全面检测BALF样本中的RNA病毒病原体,其灵敏度和基因组覆盖率与Illumina测序相当。这种快速测序平台可能更有利于在临床环境中检测RNA病毒。
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来源期刊
Journal of clinical virology plus
Journal of clinical virology plus Infectious Diseases
CiteScore
2.20
自引率
0.00%
发文量
0
审稿时长
66 days
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