LAMP-Seq: Sensitive, Scalable, and Multiplexable COVID-19 Diagnostics Using Molecular Barcoding

Autor: 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, 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.
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Nat Biotechnol
Popis: Frequent testing of large population groups combined with contact tracing and isolation measures will be crucial for containing COVID-19 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/µl at 95% confidence, and near-perfect specificity for SARS-CoV-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 Ct values up to 33, and a specificity of 99.7%, at material cost of 2.73 USD per sample. With a time-to-result less than 24 hours, 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.
Databáze: OpenAIRE