Probe-free detection method for virus RNA

Abstract

The invention discloses a probe-free detection method for virus RNA. The probe-free detection method comprises the following steps: step 1, incubating a patient sample by using an RT-PCR reagent and an RT-PCR template; step 2, separating a PCR product from a product obtained by incubation through a dielectrophoresis effect DEP; and step 3, detecting the obtained PCR product by adopting ultraviolet absorption. Probes and fluorescent dyeing are not needed, and the cost is reduced by 50% or above. Detection signals are increased based on sample-free positive detection, concentrated signals and low background signals. In addition, the incubation time of the RT-PCR reaction is shortened due to high sensitivity, and the detection speed is far higher than that of the existing probe detection method. Ultraviolet absorption is adopted for detection, so that the detection cost is reduced.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to a probe-free detection method for viral RNA. Background technique [0002] New, deadly, and highly endemic viruses occur more frequently than in the last century, and RT-PCR is currently one of the most established methods for detecting viral RNA. For example, during the outbreak of the new coronavirus, the fluorescent probe detection method of RT-PCR is an important method for quickly screening people who may be infected. This fluorescent probe detection technology can provide test results for millions of people within 6 hours, and the accuracy has reached 95%. The process is as figure 1 As shown, the collected patient samples (i.e. nasal swabs) were first treated at 37°C (>4 hours) with RT-PCR reagents and RT-PCR templates (for SARS-Cov-2 RNA) with probes (fluorescent dyes). samples taken from swabs or throat swabs) for incubation. During this process, the RT-P...

AI Technical Summary

Problems solved by technology

However, there are corresponding defects in this method, due to the dye itself, the dye attachment procedure and purification, the cost of the RT-PCR template (fluorescent dye) combined with the probe is greater than 50%, the reaction time is long (> 4 hours), in addition due to Negative detection, dilution signal and high background signal from sample, detection signal is defective

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