Details

Autor(en) / Beteiligte

• Ludwig, Kerstin U.
• Schmithausen, Ricarda M.
• Li, David
• Jacobs, Max L.
• Hollstein, Ronja
• Blumenstock, Katja
• Liebing, Jana
• Słabicki, Mikołaj
• Ben-Shmuel, Amir
• Israeli, Ofir
• Weiss, Shay
• Ebert, Thomas S.
• Paran, Nir
• Rüdiger, Wibke
• Wilbring, Gero
• Feldman, David
• Lippke, Bärbel
• Ishorst, Nina
• Hochfeld, Lara M.
• Beins, Eva C.
• Kaltheuner, Ines H.
• Schmitz, Maximilian
• Wöhler, Aliona
• Döhla, Manuel
• Sib, Esther
• Jentzsch, Marius
• Moench, Eva-Maria C.
• Borrajo, Jacob D.
• Strecker, Jonathan
• Reinhardt, Julia
• Cleary, Brian
• Geyer, Matthias
• Hölzel, Michael
• Macrae, Rhiannon
• Nöthen, Markus M.
• Hoffmann, Per
• Exner, Martin
• Regev, Aviv
• Zhang, Feng
• Schmid-Burgk, Jonathan L.

Titel

LAMP-Seq enables sensitive, multiplexed COVID-19 diagnostics using molecular barcoding

Ist Teil von

• Nature biotechnology, 2021-12, Vol.39 (12), p.1556-1562

Ort / Verlag

New York: Nature Publishing Group US

Erscheinungsjahr

2021

Quelle

MEDLINE

Beschreibungen/Notizen

• Frequent testing of large population groups combined with contact tracing and isolation measures will be crucial for containing Coronavirus Disease 2019 outbreaks. Here we present LAMP-Seq, a modified, highly scalable reverse transcription loop-mediated isothermal amplification (RT–LAMP) method. Unpurified biosamples are barcoded and amplified in a single heat step, and pooled products are analyzed en masse by sequencing. Using commercial reagents, LAMP-Seq has a limit of detection of ~2.2 molecules per µl at 95% confidence and near-perfect specificity for severe acute respiratory syndrome coronavirus 2 given its sequence readout. Clinical validation of an open-source protocol with 676 swab samples, 98 of which were deemed positive by standard RT–qPCR, demonstrated 100% sensitivity in individuals with cycle threshold values of up to 33 and a specificity of 99.7%, at a very low material cost. With a time-to-result of fewer than 24 h, low cost and little new infrastructure requirement, LAMP-Seq can be readily deployed for frequent testing as part of an integrated public health surveillance program. SARS-CoV-2 detection with LAMP-Seq combines isothermal amplification and barcoding with high-throughput sequencing.