COVID19 Multiplex qPCR kit
The assay utilises a unique two target strategy for an effective detection of all known SARS-CoV-2
strains. E-gene (FAM), S-gene (ROX) & RNaseP (HEX)
Quick Guide COVITECH kit has been designed and manufactured based on the most up-to-date and reliable sequence information. The kit protocol is based on WHO (World Health Organization) recommended approaches using the best technologies at hand. This kit targets two regions of the virus, namely E (FAM) and S (ROX). RNaseP is also included in the same tube as an endogenous control in a3-way MutiPlex RT-qPCR reaction. The positive control template supplied with the kit is packaged at a known copy number to generate a standard curve. The template can be used in single dilution where quantitative analysis of the samples is not required. The efficiency, precision and sensitivity of the kit has been investigated using numerous panels from the actual samples obtained from Iranian patients, and the kit has been optimized based upon the attained results.
The effciency of this kit has been evaluated and confrmed by QuantStudio TM 5, Rotor Gene and Applied Biosystems TM 7500 devices.
Kit Utilization Strategy:
According to the results obtained from the Iranian samples, WHO protocols, and the advised and endorsed instructions of the Quick Response Team of the Iranian Pasteur Institute, it is advised that for the purpose of preliminary screening and diagnostics, the E gene is given preference as the basis for diagnosis, and in the case of a positive result, the S gene is interpreted as the confirmation test of the SARS-Cov-2. To determine the efficiency of the amplification process and / or to quantify the virus in the sample, you can use the positive control included in the kit. The positive control is packaged at 200000 copies per microliter. A serial dilution to generate a 6-point standard curve ranging from 1million to 10 copies is recommended for quantitative analysis. Use 5ul of the positive control template in the qPCR reaction. Note: The DNA positive control template should be handled with extreme care to avoid cross over contamination. In all of the tests, make sure to use two RNA samples extracted from healthy people as the negative control of the healthy person and also as the negative control for the No Template Control ( NTC). Investigation of the kit on Iranian samples has revealed that the Ct of the E gene generally ranges between 14 to 36. Ct value of the S gene is expected to be 1 Ct higher than E gene. Positive test result is valid and acceptable if the negative controls remain negative without any amplification signal from the endogenous control (RNase P) Negative test results should be confirmed with a reliable endogenous control (RNaseP) signal.