TH18684Y - Primers for detecting the response regulator VraR (vraR) gene in methicillin-resistant Staphylococcus aureus using real-time PCR. - Google Patents

Abstract

This invention is a primer for detecting the response regulator VraR (vroR) gene in methicillin-resistant Staphylococcus aureus using real-time PCR. It consists of two primers capable of specifically binding to the vraR gene sequence on the target DNA. The primers are combined with the test reagent and a sample of genetic material, or complementary DNA, from RNA extracted from the bacteria. The genetic material is then amplified using real-time PCR. This invention can be used for both qualitative and quantitative analysis of gene expression, providing accurate, reliable, highly sensitive, and specific results that can be reported rapidly.

Claims (2)

1. Primers for detecting the response regulator VraR (vraR) gene in methicillin-resistant Staphylococcus aureus using real-time PCR. These primers consist of two strands of the VraR (vraR) gene with the following base sequences: Forward primer: 5’-TTA TCT ATC AAC GCA AAG TG -3’; Reverse primer: 5’-AAT CTG GCT TCA ACT CAT -3’. 2. Primers for detecting the response regulator VraR (vraR) gene in methicillin-resistant Staphylococcus aureus using real-time PCR, as per Patent 1, with program conditions and temperature for optimal genetic material amplification for the primer pair. The temperature program is set as follows: Step 1: Pre-denature program at 95°C for 10 minutes. Step 2: Amplification program consisting of three steps: denaturation at 95°C for 20 seconds, annealing at 53°C for 20 seconds, and extension at 72°C for 30 seconds. Fluorescence signal is measured. (fluorescence signal) once, repeated for a total of 45 cycles, with a temperature transition rate of 20 degrees Celsius per second. Step 3: Melting curve program, consisting of 3 steps: increasing the temperature to 95 degrees Celsius for 15 seconds, then decreasing the temperature to 60 degrees Celsius for 60 seconds using a temperature transition rate of 20 degrees Celsius per second, and then increasing the temperature back to 95 degrees Celsius for 15 seconds using a temperature transition rate of 0.1 degrees Celsius per second, while continuously monitoring the fluorescence signal. Step 4: Cooling program, reducing the temperature to 40 degrees Celsius for 30 seconds using a temperature transition rate of 20 degrees Celsius per second.