Intend to Use: One Step RT-qPCR kit for the detection of SARS CoV2 Virus

Technology: Multiplex gene detection and amplification assay technology

The current pandemic of a novel COVID-19 in the global human population has caused dramatic and unprecedented impact of the economy and prompted deployment of public health authorities around the world to counter the rapid spread of the virus by fasten the rapid test using diagnostic kits and vaccination process. The COVIBOND-007 is a COVID-19 RT-PCR diagnostic kit is a indigenously produced multiplex genes real-time RT-PCR test intended for the qualitative and quantitative detection of the presence of nucleic acid from the SARS-CoV-2 in nasopharyngeal swab specimens collected from individuals who meet SARS-CoV-2 clinical criteria. The approach is based on the RT-PCR method which uses three viral genes (Spike gene, Nucleocapsid gene and RNA dependent RNA polymerase gene present in open reading frame-1 region) sets of gene-specific primers and corresponding fluorescent probes to detect three specific regions within the novel coronavirus (SARS-CoV-2). This RT-PCR panel detects SARS-CoV-2 Ribonucleic acid (RNA) specifically. view all

The approach does not generate any false positives to other coronaviruses or human micro flora. The kit also contains a primer-probe set which detects human housekeeping gene, ribonuclease Protein (RNase P). That is, the Ribonuclease Protein (RNase P) serves as an internal reference control to monitor sample collection, ribonucleic acid (RNA) extraction, and amplification. It is a multiplex RT-PCR diagnostic kit, enabling quick and precise diagnosis of COVID-19 caused by SARS-CoV-2 infection.

This One Step COVIBOND 007 RT-PCR Kit (efficient Real Time) is designed for one step, real-time reverse transcription PCR (RT-PCR) using probe detection. With this kit, all RT-PCR assay steps can be performed in a single tube. Therefore, it is not necessary to add additional reagents during the reaction. So it will minimize the risk of contamination during experimental workflow. Amplified samples are monitored in real time without need for gel electrophoresis after PCR.

This kit is suitable for detection of small quantity of RNA. This kit uses imported enzymes from Japan and designed the using HPLC purified Primers manufactured in reputed research Laboratory in Europe which allows efficient and rapid cDNA synthesis, and Taq Polymerases, to enable highly efficient RT-PCR amplifications.

This RT-PCR Kit (efficient Real Time) allows cDNA synthesis from Viral RNA using Reverse Transcriptase enzyme followed by PCR amplification by Taq Polymerase enzyme in one uninterrupted PCR procedure. PCR amplification products are detected and monitored in real time with probes.

Components (for 100 reactions; 20 μl / reaction system)

1. One Step RT-PCR Master Mix contains RTase and Taq polymerase enzymes, dNTP Mixture, Mg2+ and RNase inhibitor. - 750 μl (7.5μl/reaction).
2. RNase Free dH2O. - 1.5 ml (3 μl/reaction).
3. Primer/Probe Assay mixture contains forward and reverse primers of S, RDRP, N &RNaseP genes along with corresponding detector probes.- 450 μl (4.5μl/reaction).
4. Positive RNA control. - 250 μl (5 μl/ reaction).
5. Recommended testing RNA sample volume- 5 μl / reaction.

COVID Sampling Requirements

• Applicable sample types: upper respiratory tract samples (including throat swabs, nasal swabs, nasopharyngeal extracts, deep cough sputum); lower respiratory tract samples (including respiratory tract extracts, bronchial lavage fluid, lung tissue biopsy samples).
• Sample collection: Collect according to the conventional sample collection method and place in VTM tubes and transport to the RTPCR laboratory.

RTPCR Amplification Set up

The recommended protocol for PCR reactions is the standard protocol described below.

• Holding Stage:Reverse transcription reaction

1. 52°C 5 min
2. 95°C 10 sec

• Cycling Stage:PCR reaction (35 cycles)

1. 95°C 5 sec
2. 60°C 30 sec

(Documents to attach: COA, Package Insert, Marketing brochure)