Journal: ACS Central Science

Article Title: Assay Techniques and Test Development for COVID-19 Diagnosis

doi: 10.1021/acscentsci.0c00501

Figure Lengend Snippet: Reverse transcription-polymerase chain reaction (RT-PCR). The RT-PCR creates a cDNA copy of a specific segment of the viral RNA, which is converted to dsDNA that is exponentially amplified.

Article Snippet: Each test cartridge contains reagents for magnetic solid-phase extraction of viral RNA, cDNA amplification, and detection combining electrowetting and GenMark’s eSensor technology.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Amplification

Journal: ACS Central Science

Article Title: Assay Techniques and Test Development for COVID-19 Diagnosis

doi: 10.1021/acscentsci.0c00501

Figure Lengend Snippet: Reverse transcription loop-mediated isothermal amplification (RT-LAMP). Step 1: At the 3′-end of the viral RNA, reverse transcriptase and BIP primer initiate conversion of RNA to cDNA. Step 2: At the same end, DNA polymerase and B3 primer continue to generate the second cDNA strand to displace and release the first cDNA strand. Step 3: The FIP primer binds to the released cDNA strand and DNA polymerase generates the complementary strand. Step 4: F3 primer binds to the 3′ end, and DNA polymerase then generates a new strand while displacing the old strand. LAMP cycling produces various sized double-stranded looped DNA structures containing alternately inverted repeats of the target sequence as detected by a DNA indicator dye. Reagents*: Primers and master mix containing reverse transcriptase, DNA polymerase with strand displacing activity, dNTPs, and buffers.

Article Snippet: Each test cartridge contains reagents for magnetic solid-phase extraction of viral RNA, cDNA amplification, and detection combining electrowetting and GenMark’s eSensor technology.

Techniques: Reverse Transcription, Amplification, Sequencing, Activity Assay

Journal: ACS Central Science

Article Title: Assay Techniques and Test Development for COVID-19 Diagnosis

doi: 10.1021/acscentsci.0c00501

Figure Lengend Snippet: Two alternative CRISPR methods for detecting viral RNA. Method A (SHERLOCK assay ): RT-RPA (recombinase polymerase amplification) converts viral RNA to dsDNA. T7 transcription generates complementary RNA from the dsDNA template. The Cas13–tracrRNA complex binds to the target sequence, which activates the general nuclease enzyme activity of Cas13 to cleave the target sequence and the fluorescent RNA reporter. Method B (DETECTR assay ): RT-RPA (recombinase polymerase amplification) converts viral RNA to dsDNA. The Cas12a–tracrRNA complex binds to the target sequence, which activates the general nuclease enzyme activity of Cas12a to cleave the target sequence and the fluorescent RNA reporter.

Article Snippet: Each test cartridge contains reagents for magnetic solid-phase extraction of viral RNA, cDNA amplification, and detection combining electrowetting and GenMark’s eSensor technology.

Techniques: CRISPR, Recombinase Polymerase Amplification, Sequencing, Activity Assay

Journal: ACS Central Science

Article Title: Assay Techniques and Test Development for COVID-19 Diagnosis

doi: 10.1021/acscentsci.0c00501

Figure Lengend Snippet: Examples of Molecular Diagnostic Tests Used to Detect Viral Genetic Material in SARS-CoV-2

Article Snippet: Each test cartridge contains reagents for magnetic solid-phase extraction of viral RNA, cDNA amplification, and detection combining electrowetting and GenMark’s eSensor technology.

Techniques: Diagnostic Assay, Amplification, Multiplex Assay, Quantitative RT-PCR, Control, CRISPR, Lateral Flow Assay, Real-time Polymerase Chain Reaction, Extraction, RNA Extraction, Luminex, Mutagenesis