Kit for detecting rhinoviruses, respiratory syncytial viruses and parainfluenza viruses by micro-fluidic chip and application method of kit

Abstract

The invention discloses a kit for detecting rhinoviruses, respiratory syncytial viruses and parainfluenza viruses by a micro-fluidic chip and an application method of the kit. The kit comprises the multi-flux microfluidic nucleic acid detection chip, a preloaded dry powder testing agent and a positive quality control. The multi-flux microfluidic nucleic acid detection chip is used for controllingflow passages actively. The preloaded dry powder testing agent comprises a primer for specific conserved sequences of the rhinoviruses, the respiratory syncytial viruses and the parainfluenza virusesand a TagMan fluorescence probe. The kit has the advantages that the micro-fluidic chip technology is applied, all operations are completed by instruments after sample loading, convenience and simplicity in operation and high speed are realized, detection can be completed within 30-60 minutes, and pollution is avoided; the TagMan fluorescence probe PCR technology is applied to detect the rhinoviruses, the respiratory syncytial viruses and the parainfluenza viruses; designed primer and probe sequences are quite conservative in genes of the rhinoviruses, the respiratory syncytial viruses and theparainfluenza viruses, high specificity is achieved, detection results can be acquired within two hours, and sensitivity can reach 10copies/microliter.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit for detecting rhinovirus, respiratory syncytial virus and parainfluenza virus based on a microfluidic chip and a use method thereof. Background technique [0002] Rhinovirus (Human Rhinovirus, HRV), Respiratory Syncytial Virus (Respiratory Syncytial Virus, RSV) and Parainfluenza Virus (Parainfluenza Virus, PIV) infection are the most common pathogens that cause acute respiratory diseases in children, accounting for the proportion of respiratory diseases in infants and young children. The proportion is very high, which poses a great threat to the health of infants and young children. Rhinoviruses mainly cause upper respiratory tract infections, while respiratory syncytial virus and parainfluenza viruses mainly cause lower respiratory tract infections. Molecular diagnostic technology can meet the needs of rapid diagnosis of HRV, RSV and PIV, and because their genetic materia...

AI Technical Summary

Problems solved by technology

[0010] Immunoassay detection reagents are aimed at the specific antibodies produced by the body caused by pathogens in the sample. There is a window period for the detection of antibodies. It is easy to miss the diagnosis in the early stage of the disease, and the sensitivity and specificity are low. The sample is easy to be contaminated and there are many interfering substances. When it is too high or too low, it is easy to cause misjudgment of the result, and repeated detection and re-examination are required to confirm the diagnosis, and the reliability of the detection is poor; the detection by culture method generally takes more than one day, and the culture effect is poor, many pathogens cannot be cultured, and the delay is the greatest. best time to treat

[0011] Although ordinary fluorescent PCR detection reagents have good sensitivity and specificity, the kits are generally composed of multiple tubes of liquid reagents, which usually need to be stored in an environment of -20°C. During operation, each tube of reagents needs to be mixed in proportion. There are high requirements for storage, transportation conditions, operation methods, etc., and it is easy to cause unreliable test results due to improper storage and operation errors

At the same time, because there are many subtypes of influenza A virus and influenza B virus, the detection kits are generally aimed at the most common subtypes of influenza A virus and influenza B virus. Sometimes due to the variation of virus strains, there may be Cases of missed detection

Since the results of the fluorescent quantitative PCR instrument detection require professionals to analyze the amplification curve, sometimes there will be human errors.

Finally, because the rise and fall speed of the fluorescent quantitative PCR instrument is slow, the detection time is generally 1 to 2 hours, which takes a long time