Preliminary optimisation of a simplified sample preparation method to permit direct detection of SARS-CoV-2 within saliva samples using reverse-transcription loop-mediated isothermal amplification (RT-LAMP).
Preliminary optimisation of a simplified sample preparation method to permit direct detection of SARS-CoV-2 within saliva samples using reverse-transcription loop-mediated isothermal amplification (RT-LAMP).
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DOI：
10.1016/j.jviromet.2020.114048
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发表时间：
2021-03
影响因子：
3.1
通讯作者：
Fowler VL
中科院分区：
文献类型：
--
作者：
Howson ELA;Kidd SP;Armson B;Goring A;Sawyer J;Cassar C;Cross D;Lewis T;Hockey J;Rivers S;Cawthraw S;Banyard A;Anderson P;Rahou S;Andreou M;Morant N;Clark D;Walsh C;Laxman S;Houghton R;Slater-Jefferies J;Costello P;Brown I;Cortes N;Godfrey KM;Fowler VL
我们描述了一个简化的样品制备方法，允许快速和直接检测唾液中的SARS-CoV-2 RNA的优化，使用逆转录环介导的等温扩增（RT-LAMP）。在RT-LAMP之前，通过在Mucolyse™中1：1稀释，然后在10%（w/v）Chelex ® 100树脂中稀释和98 °C加热步骤2分钟来处理唾液样品，使得能够检测阳性唾液样品中的SARS-CoV-2 RNA。使用RT-LAMP，SARS-CoV-2 RNA在05：43分钟内被检测到，在3097份实时逆转录PCR（rRT-PCR）阴性唾液样本中未检测到扩增，这些样本来自服务评估研究中测试的工作人员或其他呼吸道病原体（n = 22）。唾液样本可以非侵入性地收集，不需要熟练的工作人员，并且可以从医疗保健和家庭环境中获得。重要的是，这种方法克服了对不同拭子的要求和验证，以及获得提取机器人和试剂的全球瓶颈，从而能够通过rRT-PCR进行分子检测。这种检测为在COVID-19大流行期间通过在人群规模进行定期SARS-CoV-2检测，结合隔离和接触者追踪，进行有效干预的公共卫生方法提供了可能性。
We describe the optimisation of a simplified sample preparation method which permits rapid and direct detection of SARS-CoV-2 RNA within saliva, using reverse-transcription loop-mediated isothermal amplification (RT-LAMP). Treatment of saliva samples prior to RT-LAMP by dilution 1:1 in Mucolyse™, followed by dilution in 10 % (w/v) Chelex© 100 Resin and a 98 °C heat step for 2 min enabled detection of SARS-CoV-2 RNA in positive saliva samples. Using RT-LAMP, SARS-CoV-2 RNA was detected in as little as 05:43 min, with no amplification detected in 3097 real-time reverse transcription PCR (rRT-PCR) negative saliva samples from staff tested within a service evaluation study, or for other respiratory pathogens tested (n = 22). Saliva samples can be collected non-invasively, without the need for skilled staff and can be obtained from both healthcare and home settings. Critically, this approach overcomes the requirement for, and validation of, different swabs and the global bottleneck in obtaining access to extraction robots and reagents to enable molecular testing by rRT-PCR. Such testing opens the possibility of public health approaches for effective intervention during the COVID-19 pandemic through regular SARS-CoV-2 testing at a population scale, combined with isolation and contact tracing.
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影响因子：
3.1
作者：
Fowler, Veronica L.;Howson, Emma L. A.;King, Donald P.
通讯作者：
King, Donald P.
通过逆转录环介导的等温扩增测定快速直观地检测 2019 新型冠状病毒 (SARS-CoV-2)
影响因子：
14.2
作者：
Yan, C.;Cui, J.;Yuan, J.
通讯作者：
Yuan, J.
DOI：
10.1016/j.jinf.2020.10.039
发表时间：
2021-01
期刊：
The Journal of infection
影响因子：
--
作者：
Fowler VL;Armson B;Gonzales JL;Wise EL;Howson ELA;Vincent-Mistiaen Z;Fouch S;Maltby CJ;Grippon S;Munro S;Jones L;Holmes T;Tillyer C;Elwell J;Sowood A;de Peyer O;Dixon S;Hatcher T;Patrick H;Laxman S;Walsh C;Andreou M;Morant N;Clark D;Moore N;Houghton R;Cortes NJ;Kidd SP
通讯作者：
Kidd SP
影响因子：
3.8
作者：
Azzi, Lorenzo;Carcano, Giulio;Baj, Andreina
通讯作者：
Baj, Andreina
影响因子：
64.8
作者：
Mulligan, Mark J.;Lyke, Kirsten E.;Jansen, Kathrin U.
通讯作者：
Jansen, Kathrin U.