ZELLX® real time qPCR SARS-CoV-2 detection and extraction kits

The novel human coronavirus, officially termed 2019-nCoV and more commonly known as COVID-19, had it first recorded cases in the Chinese city of Wuhan in late 2019. However, the coronavirus is strongly linked to a strain then labelled RaBtCoV/4991 that had caused multiple deaths among miners clearing bat faeces from an abandoned copper mine near the town of Tongguuan in the Mojian region of Southern China during 2012.

The 2019-nCoV causes pneumonia and is genetically different from other common human coronaviruses such as 229E, NL63, OC43 and HKU1 that cause seasonal acute respiratory illnesses. It is also genetically distinct from the two newer human coronaviruses, MERS-CoV and SARS-CoV.

The resulting global coronavirus pandemic that spread outwards from Wuhan has created intense demand for real-time SARS-CoV-2 detection and rapid genetic extraction from samples. Accordingly, ZELLX® has introduced three new diagnostic and analytical kits that use quantitative polymerase chain reaction (qPCR) technology.

ZELLX® SARS‐CoV-2 Real Time-qPCR Detection Kits

The ZELLX® SARS-CoV-2 RT-qPCR assay design is based on US Center for Disease Control (CDC) guidelines for SARS-CoV-2 real-time PCR detection and allows specific detection of SARS-CoV-2 in less than two hours using real time RT-PCR in respiratory specimens, including sputum; nasopharyngeal, oropharyngeal aspirates, washes or swabs, and tracheal aspirates.

The ZELLX® RT-qPCR Kit allows efficient cDNA synthesis and Real-Time PCR in a single tube. The qPCR Master Mix provided in the kit contains all the reagent (except PCR primers, probe and template) needed for running PCR reactions. To prevent the degradation of viral RNA by RNases, the kit comes with a vial of Ribonuclease (RNAse) inhibitor.

The kit also contains one set of primers and fluorescent probe designed according to target region of the virus nucleocapsid (N) gene and to detect the human RNase P used as a control to assess specimen quality. The probes have readout in different channels. Coronavirus SARS-CoV-2 RNA targets are amplified and detected in the FAM channel, while human RNase P RNA targets are amplified and detected in the HEX, VIC or JOE channel (depending on the equipment).

The kit is offered in two versions:

• ZX-22100-100: SARS‐CoV-2 Real Time-qPCR Detection Kit for 100 test reactions (RXNS)
• ZX-22102-250: SARS-CoV-2 RT-qPCR Detection kit (N&RP) for 250 RXNS

The 100 test version does not include a positive control (PC), needed to demonstrate that the PCR amplification is working efficiently with the supplied components. To confirm absence of contamination, a non-template (negative) control reaction should be included every time the kit is used.

The 250-test kit includes a separate Reverse Transcriptase.

Both kits allow efficient cDNA synthesis and Real-Time PCR in a single tube, including a qPCR Master Mix that contains all reagents needed (except PCR primers, probe and template) to run PCR reactions.

The tests will remain stable in storage at -20°C for up to two years.

Viral RNA Extraction Kit

The ZELLX® ZX-22101-500 Viral RNA Extraction Kit is designed for the quick purification of viral RNA from cell-free samples such as serum, plasma, urine, and other body fluids, cell culture supernatants and rinse liquid from swabs samples.

For safety, the kit uses a non-phenol-chloroform extraction method in which viral RNA molecules bind to the silica-based membrane, and impurities such as proteins and nucleases are removed by thorough washing with Wash Buffer. The RNA is then eluted in sterile, RNase free water.

The isolated viral RNA is ready to use and should be stored at -70°C. The procedure can be used for isolation of viral RNA from a broad range of viruses. However, performance cannot be guaranteed for every virus species and must be separately validated.

The kit exhibits 100% specificity and sensitivity of 98.3% with quantity of purified viral RNA extracted depending on various parameters including sample type, virus titer, sample source, transport, storage, and age. The Kit additionally contains carrier RNA that improves binding and recovery of low-concentrated viral RNA.

The Extraction Kit provides sufficient materials for 500 test reactions and can be used in RT-PCR and qRT-PCR for viral detection, viral load monitoring, and viral genotyping.

Resources

Click on ZX-22100-100 (100 RXNS) | Coronavirus (SARS‐CoV-2) Real Time-qPCR Detection Kit for product details.
Click on ZX-22102-250 (250 RXNS) | SARS-CoV-2 – RT-qPCR Detection kit (N&RP) for product details.