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首页> 外文期刊>Journal of Clinical Microbiology >Development, Technical Performance, and Clinical Evaluation of a NucliSens Basic Kit Application for Detection of Enterovirus RNA in Cerebrospinal Fluid
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Development, Technical Performance, and Clinical Evaluation of a NucliSens Basic Kit Application for Detection of Enterovirus RNA in Cerebrospinal Fluid

机译：NucliSens基本试剂盒用于检测脑脊液中肠病毒RNA的开发，技术性能和临床评估

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The combination of nucleic acid sequence-based amplification and electrochemiluminescence detection was used to develop an internally controlled, highly sensitive and specific assay for the detection of enterovirus (EV) RNA in cerebrospinal fluid (CSF). The analytical performance of the assay was determined using both in vitro-transcribed EV RNAs and viral culture isolates. The sensitivity of the assay was 10 EV RNA copies per amplification reaction. The assay detected all enteroviral isolates tested with no cross-reactivity to 21 nonenteroviral species, including rhinovirus and parechovirus. The clinical performance of the assay was evaluated by testing 992 CSF specimens collected from adult and pediatric patients. NucliSens EV results from a subset of 327 CSF samples were compared to viral culture of nasopharyngeal specimens and rectal swabs (n = 195) and/or CSF (n = 212). Of the 212 CSF samples, 96 samples were positive by either the NucliSens EV assay (94/96; 97.9%) or culture (63/96; 65.6%), and 61/96 (63.5%) were positive by both methods. The inclusion of an EV-specific internal control monitored the entire process, including the efficiency of nucleic acid extraction, amplification, and detection. In total, only five blood-clotted CSF samples (0.5%) were inhibited. The NucliSens EV assay demonstrated superior sensitivity over viral culture (P < 0.001), excellent specificity, clear delineation of positive samples, and minimal amplification inhibition.
机译：基于核酸序列的扩增和电化学发光检测相结合，可用于开发一种内部控制的，高灵敏度和特异性的检测方法，用于检测脑脊液（CSF）中的肠病毒（EV）RNA。使用体外转录的EV RNA和病毒培养分离物测定测定的分析性能。该测定法的灵敏度是每个扩增反应10个EV RNA拷贝。该检测方法检测到所有与21种非肠道病毒无交叉反应的肠道病毒分离株，包括鼻病毒和副病毒。通过测试从成人和儿科患者收集的992例CSF标本，评估了该测定法的临床性能。将来自327个CSF样本的NucliSens EV结果与鼻咽标本和直肠拭子（ n = 195）和/或CSF（ n = 212）的病毒培养进行了比较。在212个CSF样本中，通过NucliSens EV分析（94/96; 97.9％）或培养（63/96; 65.6％）呈阳性的96个样本，两种方法均为61/96（63.5％）呈阳性。包括EV专用内部控制可监控整个过程，包括核酸提取，扩增和检测的效率。总共仅抑制了五个血凝的CSF样品（0.5％）。 NucliSens EV测定法显示出优于病毒培养物的灵敏度（ P <0.001），出色的特异性，清晰的阳性样品轮廓以及最小的扩增抑制作用。

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