【 以下文字转载自 Military 讨论区 】 发信人: lubbock12 (非老非小将), 信区: Military 标 题: 诶呀，季博士把德国人创造激情也点燃了 发信站: BBS 未名空间站 (Tue Aug 18 13:27:53 2020, 美东)
也开始沉溺于lamp变色检测而不能自拔
SARS-CoV-2 RNA Extraction Using Magnetic Beads for Rapid Large-Scale Testing by RT-qPCR and RT-LAMP
Rapid large-scale testing is essential for controlling the ongoing pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The standard diagnostic pipeline for testing SARS-CoV-2 presence in patients with an ongoing infection is predominantly based on pharyngeal swabs, from which the viral RNA is extracted using commercial kits, followed by reverse transcription and quantitative PCR detection. As a result of the large demand for testing, commercial RNA extraction kits may be limited and, alternatively, non-commercial protocols are needed. Here, we provide a magnetic bead RNA extraction protocol that is predominantly based on in- house made reagents and is performed in 96-well plates supporting large- scale testing. Magnetic bead RNA extraction was benchmarked against the commercial QIAcube extraction platform. Comparable viral RNA detection sensitivity and specificity were obtained by fluorescent and colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) using a primer set targeting the N gene, as well as RT-qPCR using a primer set targeting the E gene, showing that the RNA extraction protocol presented here can be combined with a variety of detection methods at high throughput. Importantly, the presented diagnostic workflow can be quickly set up in a laboratory without access to an automated pipetting robot. View Full-Text https://www.mdpi.com/1999-4915/12/8/863
【 以下文字转载自 Military 讨论区 】
发信人: lubbock12 (非老非小将), 信区: Military
标 题: 诶呀，季博士把德国人创造激情也点燃了
发信站: BBS 未名空间站 (Tue Aug 18 13:27:53 2020, 美东)
也开始沉溺于lamp变色检测而不能自拔
SARS-CoV-2 RNA Extraction Using Magnetic Beads for Rapid Large-Scale Testing by RT-qPCR and RT-LAMP
Rapid large-scale testing is essential for controlling the ongoing pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The
standard diagnostic pipeline for testing SARS-CoV-2 presence in patients
with an ongoing infection is predominantly based on pharyngeal swabs, from
which the viral RNA is extracted using commercial kits, followed by reverse transcription and quantitative PCR detection. As a result of the large
demand for testing, commercial RNA extraction kits may be limited and,
alternatively, non-commercial protocols are needed. Here, we provide a
magnetic bead RNA extraction protocol that is predominantly based on in-
house made reagents and is performed in 96-well plates supporting large-
scale testing. Magnetic bead RNA extraction was benchmarked against the
commercial QIAcube extraction platform. Comparable viral RNA detection
sensitivity and specificity were obtained by fluorescent and colorimetric
reverse transcription loop-mediated isothermal amplification (RT-LAMP) using a primer set targeting the N gene, as well as RT-qPCR using a primer set
targeting the E gene, showing that the RNA extraction protocol presented
here can be combined with a variety of detection methods at high throughput. Importantly, the presented diagnostic workflow can be quickly set up in a
laboratory without access to an automated pipetting robot. View Full-Text
https://www.mdpi.com/1999-4915/12/8/863