RNA transcriptional biosignature analysis for identifying febrile infants with serious bacterial infections in the emergency department: a feasibility study.
RNA transcriptional biosignature analysis for identifying febrile infants with serious bacterial infections in the emergency department: a feasibility study.
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DOI：
10.1097/pec.0000000000000324
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发表时间：
2015-01
影响因子：
1.4
通讯作者：
Febrile Infant Working Group for the Pediatric Emergency Care Applied Research Network (PECARN)
中科院分区：
文献类型：
--
作者：
Mahajan P;Kuppermann N;Suarez N;Mejias A;Casper C;Dean JM;Ramilo O;Febrile Infant Working Group for the Pediatric Emergency Care Applied Research Network (PECARN)
在多中心儿科急诊研究网络中，发展基础设施并证明进行基于微阵列的RNA转录谱分析以诊断60天及以下发热婴儿严重细菌感染的可行性。我们设计了一项前瞻性多中心队列研究，目的是纳入4000多名60天及以下的发热婴儿。为了确保在急诊科（ED）环境中进行复杂基因组研究的成功，我们在包括21个站点的儿科急诊应用研究网络中建立了一个基础设施，以评估年幼发热婴儿的RNA转录谱。我们制定了一份全面的操作手册，并培训现场调查员获取和处理血液样本进行RNA提取和基因组分析。我们制定了血液样本采集、处理、储存、运输和分析的标准操作程序。我们计划从符合条件的患者中前瞻性地识别、招募和收集1ml血液样本进行基因组分析，以确定研究程序中的后勤问题。最后，我们计划在中央微阵列实验室批量采集血样，测定RNA的数量和质量，并与儿科急诊应用研究网络数据协调中心组织数据分析。下面我们就基础设施的建立和第一年基于有限数量患者的可行性成功进行报告。我们成功地在21个急诊科建立了基础设施。在前5个月，我们招募了79%（94名中的74名）符合条件的发热婴儿。我们能够从74%的参与者（74人中有55人）获得并运送1ml血液，每个参与ED至少有1个样本。55个样本被运送到微阵列实验室进行评估，95%（55人中有52人）的血液样本质量足够，含有足够的RNA用于表达分析。在多中心儿科急诊科研究背景下，有可能建立一个强大的基础设施，对年幼的发热婴儿进行基因组研究。所获得的足够数量和高质量的RNA表明，在这种情况下，可以成功地进行全血转录谱分析，用于年幼发热婴儿的诊断评估。
To develop the infrastructure and demonstrate the feasibility of conducting microarray-based RNA transcriptional profile analyses for the diagnosis of serious bacterial infections in febrile infants 60 days and younger in a multicenter pediatric emergency research network. We designed a prospective multicenter cohort study with the aim of enrolling more than 4000 febrile infants 60 days and younger. To ensure success of conducting complex genomic studies in emergency department (ED) settings, we established an infrastructure within the Pediatric Emergency Care Applied Research Network, including 21 sites, to evaluate RNA transcriptional profiles in young febrile infants. We developed a comprehensive manual of operations and trained site investigators to obtain and process blood samples for RNA extraction and genomic analyses. We created standard operating procedures for blood sample collection, processing, storage, shipping, and analyses. We planned to prospectively identify, enroll, and collect 1 mL blood samples for genomic analyses from eligible patients to identify logistical issues with study procedures. Finally, we planned to batch blood samples and determined RNA quantity and quality at the central microarray laboratory and organized data analysis with the Pediatric Emergency Care Applied Research Network data coordinating center. Below we report on establishment of the infrastructure and the feasibility success in the first year based on the enrollment of a limited number of patients. We successfully established the infrastructure at 21 EDs. Over the first 5 months we enrolled 79% (74 of 94) of eligible febrile infants. We were able to obtain and ship 1 mL of blood from 74% (55 of 74) of enrolled participants, with at least 1 sample per participating ED. The 55 samples were shipped and evaluated at the microarray laboratory, and 95% (52 of 55) of blood samples were of adequate quality and contained sufficient RNA for expression analysis. It is possible to create a robust infrastructure to conduct genomic studies in young febrile infants in the context of a multicenter pediatric ED research setting. The sufficient quantity and high quality of RNA obtained suggests that whole blood transcriptional profile analysis for the diagnostic evaluation of young febrile infants can be successfully performed in this setting.