LAMP assays have a variety of readouts due to the large amount of DNA generated, including fluorescence using an intercalating DNA dye, turbidity, or a drop in the pH if the reaction is minimally buffered ( 1, 4, 5). Furthermore, although the strand-displacing polymerase has reverse transcriptase activity, a reverse transcriptase can be included to improve sensitivity within the reaction when detecting an RNA target (RT-LAMP), such as the SARS-CoV-2 RNA. Such reactions generate microgram quantities of DNA in a very short period of time at a single reaction temperature. Utilizing a strand-displacing polymerase and loops formed during this reaction, a fast amplification reaction occurs upon proper oligo binding to the desired target. LAMP is a method of isothermal DNA replication that utilizes, in an accelerated format, six DNA oligos that hybridize with eight different regions of a target molecule ( 2, 3). It should increase the availability of SARS-CoV-2 testing as well as expand the settings in which this testing can be performed. This simple inactivation and purification pipeline is inexpensive and compatible with other downstream RNA detection platforms and uses readily available reagents. The inactivation and purification protocol, combined with the RT-LAMP assay, brings the sensitivity to at least 1 viral RNA copy per microliter in a sample. To further increase the sensitivity, a purification protocol compatible with this inactivation method was developed. This protocol, combined with the RT-LAMP assay, has a sensitivity of at least 50 viral RNA copies per microliter in a sample. A rapid inactivation protocol capable of inactivating virions, as well as endogenous nucleases, was optimized to increase sensitivity and sample stability. To address this need for increased testing, we have developed a sensitive reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay compatible with current reagents, which utilizes a colorimetric readout in as little as 30 min. The effects will continue to grow and worsen if economies begin to open without the proper precautions, including expanded diagnostic capabilities. Sweetwater.The current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has had an enormous impact on society worldwide, threatening the lives and livelihoods of many.Arturia (opens in new tab) - all the current deals in one place.
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