SARS-COV-2 RT-qPCR Assay (Triple Fluorescence RT-PCR Method), detection of ORF1ab and N genes of the SARS CoV 2

The SARS-COV-2 RT qPCR Assay is a real time (rt) reverse transcriptase (RT) polymerase chain reaction (PCR) test intended for the qualitative detection of ORF1ab and N genes of the SARS CoV 2 in nasopharyngeal ( NP) swab, oropharyngeal (OP) swab and sputum specimen collected by a healthcare worker, from patients suspected of COVID-19 by their health care provider.

Features

• High Accuracy
• Limit of Detection: 200 copies/mL
• Sensitivity: 100 %
• Specificity: 99.1%
• Accuracy: 99.3%

Convenient Operation

Triplex-detections in a single tube (ORF1ab & N genes, human RNAse P (RNP) gene) Primer and probe premixed liquid form

Reliable

• Anti-pollution system: UDG
• Internal control: Human RNaseP (RNP) gene
• Positive control: Pseudovirus

Fast

Result in about 1 hour

APPLICABLE INSTRUMENTS:

Real-time PCR instrument with FAM, TEXAS RED/ROX and HEX/VIC channels, such as ABI7500, ABI Q3, ABI Q6, Bio Rad CFX96.

Novel coronavirus pneumonia (coronavirusdisease2019, covid-19). sars-cov-2 infection is a serious global public health problem that has caused severe economic losses worldwide, leading the World Health Organization (WHO) to characterize the sars-cov-2 outbreak as a public health emergency of international concern.

Early screening of suspected cases and isolation and treatment of patients infected with sars-cov-2 are key to controlling the outbreak, as there is no specific drug or vaccine available to treat neocrown pneumonia. Currently, the gold standard for the diagnosis of sars-cov-2 infection is the reversetranscription-polymerase chain reaction (rt-pcr). Since the release of the complete genome of sars-cov-2, public health laboratories, clinical laboratories and diagnostic companies from different countries have developed a variety of rt-pcr methods for sars-cov-2 detection, targeting different genes and genomic regions of the viral genome (orf1ab, rdrp, n, e, etc.).

However, the rt-pcr method has revealed many drawbacks and shortcomings in practice, especially during public health emergencies such as the current sars-cov-2 outbreak. rt-pcr methods are cumbersome to perform and need to be performed on pcr instruments by trained professional laboratory technicians.

This makes it difficult to meet the demand for on-site testing, and for remote areas or primary hospitals with limited resources, samples must be transported to a higher level hospital with rt-pcr testing facilities for testing. This is not only time consuming but also increases the risk of virus exposure. In addition, rt-pcr for sars-cov-2 testing during the covid-19 pandemic produced a high false negative rate (30-50%) for a number of reasons, which remains a non-negligible problem.

The catalytic hairpin dna self-assembly reaction (catalytichairpinassembly, cha) is an isothermal enzyme-free nucleic acid signal amplification system. Two sets of complementary single-stranded dna probes with a hairpin structure are designed based on the target rna fragment being detected. When the target fragment is not present, both sets of single-stranded dna are present in a hairpin structure.

When the target rna fragment is present in the system, the two sets of single-stranded dna probes with hairpin structure will open the hairpin structure in turn and form a dna double strand under the catalytic effect of the target rna. The whole catalytic process does not consume the target rna fragment, and the target rna fragment urges the two sets of single-stranded dna probes to form a double strand before continuing to the next round of reaction, thus creating signal amplification. Finally, the detection of the target rna fragment is achieved by detecting the double-stranded dna probe.