Development of a New Multiplex Real-Time RT-PCR Assay for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Detection.
Development of a New Multiplex Real-Time RT-PCR Assay for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Detection.
复制标题
DOI：
10.1016/j.jmoldx.2020.09.004
复制
发表时间：
2020-12
期刊：
影响因子：
--
通讯作者：
Berry GJ
中科院分区：
文献类型：
--
作者：
Zhen W;Berry GJ
本研究描述了一种新的多重实时RT-PCR检测严重急性呼吸综合征冠状病毒2型（SARS-CoV-2），设计的引物扩增刺突表面糖蛋白（S基因）上的108 bp的目标和水解TaqMan探针设计，特异性检测SARS-CoV-2。评价了该新检测方法的检测限（LOD）和临床性能。使用灭活病毒进行的LOD研究显示，性能与改良CDC检测试剂盒相当，Northwell Health Laboratories实验室开发检测试剂盒（NWHL LDT）的最终LOD为1301 ± 13基因组当量/mL，而改良CDC检测试剂盒的最终LOD为1249 ± 14基因组当量/mL。此外，270份鼻咽拭子样本的临床评价显示，与改良CDC检测相比，阳性一致率为98.5%，阴性一致率为99.3%。NWHL LDT多重设计允许每个平板检测91名患者，而改良CDC检测试剂盒最多每个平板检测29名患者，在这两个参数都很关键的时候，每次运行可以检测更多患者，节省试剂。结果表明，NWHL LDT多重检测执行以及修改的CDC检测，但更有效和成本效益，可用作诊断检测和流行病学监测和临床管理的SARS-CoV-2。
This research describes the development of a new multiplex real-time RT-PCR test for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with primers designed to amplify a 108 bp target on the spike surface glycoprotein (S gene) and a hydrolysis TaqMan probe designed to specifically detect SARS-CoV-2. The limit of detection (LOD) and clinical performance of this new assay were evaluated. A LOD study with inactivated virus exhibited performance equal to the modified CDC assay, with a final LOD of 1301 ± 13 genome equivalents/mL for the Northwell Health Laboratories laboratory-developed test (NWHL LDT) versus 1249 ± 14 genome equivalents/mL for the modified CDC assay. In addition, a clinical evaluation with 270 nasopharyngeal swab specimens exhibited 98.5% positive percent agreement and 99.3% negative percent agreement compared with the modified CDC assay. The NWHL LDT multiplex design allows testing of 91 patients per plate, versus a maximum of 29 patients per plate on the modified CDC assay, providing the benefit of testing significantly more patients per run and saving reagents, during a time when both of these parameters are critical. The results show that the NWHL LDT multiplex assay performs as well as the modified CDC assay but is more efficient and cost-effective and can be used as a diagnostic assay and for epidemiologic surveillance and clinical management of SARS-CoV-2.
登录
查看更多内容
影响因子：
--
作者：
Corman VM;Muth D;Niemeyer D;Drosten C
通讯作者：
Drosten C
影响因子：
8.8
作者：
Uhteg, Katharine;Jarrett, Junko;Mostafa, Heba H.
通讯作者：
Mostafa, Heba H.
影响因子：
1.9
作者：
LANDIS, JR;KOCH, GG
通讯作者：
KOCH, GG
与可能源自蝙蝠的新型冠状病毒相关的肺炎爆发
影响因子：
64.8
作者：
Zhou, Peng;Yang, Xing-Lou;Shi, Zheng-Li
通讯作者：
Shi, Zheng-Li
影响因子：
9.4
作者：
Zhen, Wei;Smith, Elizabeth;Berry, Gregory J.
通讯作者：
Berry, Gregory J.