Kit for detecting three respiratory pathogens based on micro-fluidic chip and use method of kit

Abstract

The invention discloses a kit for detecting respiratory adenoviruses, streptococcus pneumoniae and bordetella pertussis based on a micro-fluidic chip and a use method of the kit. The kit comprises a multi-flux micro-fluidic nucleic acid detection chip for actively controlling flow paths, a pre-filled dry powder detection reagent and a positive quality control product, wherein the pre-filled dry powder detection reagent contains primers with specific conserved sequences for the respiratory adenoviruses, the streptococcus pneumoniae and the bordetella pertussis and a TaqMan fluorescent probe. The kit disclosed by the invention adopts a micro-fluidic chip technology, all operation processes are completed by instruments after application of a sample, the operation is simple and convenient, thespeed is high, the detection can be completed within 30-60 min, and pollution can be avoided; the fluorescent PCR technology of the TaqMan probe is adopted to detect the respiratory adenoviruses, thestreptococcus pneumoniae and the bordetella pertussis; the sequences of the designed primers and probe are very conserved in genes of the respiratory adenoviruses, the streptococcus pneumoniae and the bordetella pertussis, and the designed primers and probe have high specificity; and detection results can be obtained within 2h, and the sensitivity can reach 10 copies / mu L.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit for detecting respiratory adenovirus, Streptococcus pneumoniae and Bacillus pertussis based on a microfluidic chip and a use method thereof. Background technique [0002] Respiratory Adenovirus (ADV), Streptococcus pneumoniae (SP) and Bordetella pertussis (Bordetella pertussis, BP) are common pathogens that cause respiratory diseases in children and pose a great threat to the health of infants and young children. ADV, SP and BP infection has become a global problem. Molecular diagnostic technology can meet the needs of rapid diagnosis of ADV, SP and BP, and because their genetic material is deoxyribonucleic acid, no reverse transcription process is required after sample extraction, and molecular detection can be carried out directly, so it can be quickly and accurately Carry out pathogen screening, so as to cooperate with correct treatment, reduce the misuse of antibacteri...

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