Comprehensive detection of viruses in pediatric patients with acute liver failure using next-generation sequencing
Comprehensive detection of viruses in pediatric patients with acute liver failure using next-generation sequencing
复制标题
DOI：
10.1016/j.jcv.2017.10.001
复制
发表时间：
2017-11-01
影响因子：
8.8
通讯作者：
Ito, Yoshinori
中科院分区：
文献类型：
--
作者：
Suzuki, Takako;Kawada, Jun-ichi;Ito, Yoshinori
背景：小儿急性肝衰竭（PALF）是一种罕见且严重的综合征，经常需要肝移植。病毒是本病最常见的病因之一，然而，许多患者的致病病毒尚未确定。近年来，新一代测序（NGS）技术被广泛应用于不明病因传染病病原体的综合检测。目的：评价基于ngs的PALF或病因不明的急性肝炎患者病原病毒检测方法。研究设计：为了检测患者血清/血浆中存在的病毒来源的DNA和RNA序列，对DNA和RNA进行测序。首先，我们验证了NGS在临床血清/血浆样本中检测病毒病原体的能力，并比较了不同的商用RNA文库制备方法。然后，使用NGS对14例PALF或病因不明的急性肝炎患者的血清/血浆进行了评估。结果：在3种RNA文库制备方法中，Ovation RNA- seq System V2对RNA病毒序列的检测灵敏度最高。在14例患者中，各有1例患者血清中检测到转矩病毒、腺相关病毒和隐身病毒的序列，但这3种病毒的病理生理作用尚不清楚。在其余11例患者中未检测到显著的病毒读数。结论：这一发现可能是由于转诊时血液中病毒滴度低或非感染性原因可能更常见。这些结果表明，基于ngs的方法有可能检测临床样本中的病毒病原体，并有助于澄清PALF的病因。
Background: Pediatric acute liver failure (PALF) is a rare and severe syndrome that frequently requires liver transplantation. Viruses are one of the most frequent causes of this disease, however, pathogenic viruses are not determined in many patients. Recently next-generation sequencing (NGS) has been applied to comprehensively detect pathogens of infectious diseases of unknown etiology.Objectives: To evaluate an NGS-based approach for detecting pathogenic viruses in patients with PALF or acute hepatitis of unknown etiology.Study design: To detect virus-derived DNA and RNA sequences existing in sera/plasma from patients, both DNA and RNA sequencing were performed. First, we validated the ability of NGS to detect viral pathogens in clinical serum/plasma samples, and compared different commercial RNA library preparation methods Then, serum/plasma of fourteen patients with PALF or acute hepatitis of unknown etiology were evaluated using NGS.Results: Among three RNA library preparation methods, Ovation RNA-Seq System V2 had the highest sensitivity to detect RNA viral sequences. Among fourteen patients, sequence reads of torque teno virus, adeno-associated virus, and stealth virus were found in the sera of one patient each, however, the pathophysiological role of these three viruses was not clarified. Significant virus reads were not detected in the remaining 11 patients.Conclusions: This finding might be due to low virus titer in blood at the time of referral or a non-infectious cause might be more frequent. These results suggest an NGS-based approach has potential to detect viral pathogens in clinical samples and would contribute to clarification of the etiology of PALF.