RT-PCR Corona Virus Test Kit
Wuhan-CoV (2019-nCoV) qPCR Detection Kit 100 Reaction
Figure.1 Targets of the oligonucleotide sets used in the kit. RdRp probe targeting only Wuhan CoV shows 100% similarity to the Wuhan CoV sequence and has at least 12% difference with the SARS CoV sequences. The oligonucleotide set targeting E gene has at least 96% similarity to all the sequences and can amplify the Wuhan CoV and SARS CoV sequences. The N gene targeted oligonucleotide set differs up to 28% with some Bat SARS CoV sequences and can amplify Wuhan CoV and some SARS CoV sequences. Details of alignments using 6 different Wuhan CoV genomes and 34 different SARS CoV genomes can be found in World Health Organization guidelines (WHO Protocol, 13.01.2020, Diagnostic detection of Wuhan coronavirus 2019 by real-time RT-PCR).
Intended Use and Test Principle: Kit is used for detecting "severe acute respiratory syndrome (SARS)-related coronavirus (SARS-CoV)" and the epidemic "2019 novel CoV (2019-nCoV) / Wuhan CoV)”. The kit is applied to nucleic acid isolates from nasopharyngeal aspirate / lavage, bronchoalveolar lavage, nasopharyngeal swab, oropharyngeal swab and sputum samples. Rapid diagnosis with the kit is achieved via onestep reverse transcription (RT) and real-time PCR (qPCR) (RT-qPCR) targeting SARS-CoV-specific E (envelope) and N (nucleocapsid) gene fragments and Wuhan-CoV-specific RdRp (RNA-dependent RNA polymerase) gene fragment.
Analytical Specifications: Kit is validated with Roche LightCycler® 96, Bio-Rad CFX96 Touch™, Qiagen Rotor-Gene® 5 Plex Real-Time PCR Systems. The kit's detection limit (LOD) is the lowest analyte concentration that can be detected with a 95% probability. The LOD of the oligonucleotide sets included in the kit, inclusivity and exclusivity studies were carried out using 35 different genotypes and published by the World Health Organization (WHO Protocol, 13.01.2020, Diagnostic detection of Wuhan coronavirus 2019 by real-time RT-PCR). LOD of the test for the E, RdRp and N genes are 5.2, 3.8 and 8.3 copy-RNA/reaction respectively. The kit is validated with RINATM M14 Nucleic Acid Extraction Robot (Cat No: RINA-M14-01) and its consumables (Cat No: RN-NA-14-111-100) and the manual Bio-Speedy® Nucleic Acid Isolation Kit (Cat No: BS-NA102-100).
Warnings: 1) The kit should be stored away from nucleic acid sources and qPCR amplicons. 2) The components in the kit should not be mixed with components with different lot numbers or chemicals of the same name but from different manufacturers. 3) Master stock reagents should be kept on the cold block during the PCR setup; if possible, the PCR setup should be performed on the cold block. 4) Kit components should be mixed by gently shaking before use. 5) The micropipettes used for pipetting qPCR mixes and template nucleic acids should be separate. 6) Template nucleic acid and positive control tubes should always be kept closed, except for fluid transfers. 7) The wipeable surfaces of the rooms, benches and devices where the analysis is performed should be cleaned regularly with 10% NaClO. 8) The qPCR completed reaction tubes should be disposed of before opening in the laboratory.