Kit for detecting plurality of pathogens on basis of microfluidic chip and using method of kit

Abstract

The invention discloses a kit for detecting a plurality of pathogens on the basis of a microfluidic chip and a using method of the kit. The kit comprises a multiflux microfluidic nucleic acid detection chip for actively controlling a flow passage, a preloaded dry powder detection reagent and a positive quality control material, wherein the preloaded dry powder detection reagent contains a primer and a Taqman fluorescent probe of a specific conserved sequence of mycoplasma pneumoniae, chlamydia pneumoniae and legionella pneumoniae. According to the kit, a microfluidic chip technology is adoptedand all operations after sample loading are completed by the apparatus, so that convenience for operation and high speed are achieved, detection can be completed within 30 to 60min and pollutions areavoided; a Taqman probe fluorescence PCR (polymerase chain reaction) technology is adopted for detection of the mycoplasma pneumoniae, the chlamydia pneumoniae and the legionella pneumoniae, the sequence of the primer and the probe is designed to be conserved in genes of the mycoplasma pneumoniae, the chlamydia pneumoniae and the legionella pneumoniae and high in specificity, a detection result can be obtained within 2 hours and the sensitivity can reach 10copies / mu L.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit for detecting Mycoplasma pneumoniae, Chlamydia pneumoniae and Legionella pneumophila based on a microfluidic chip and a use method thereof. Background technique [0002] Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP) and Legionella pneumophila (Legionella pneumoniae, LP) infections are important infectious diseases in the world, which can occur in children of all ages, and respiratory tract infections are the most common , the infection site can run through the entire respiratory tract, and has certain infectivity. MP, CP, and LP are common pathogenic bacteria in children with acute upper respiratory tract infection, and they are also the main pathogenic bacteria that cause community-acquired pneumonia. Mycoplasma pneumoniae pneumonia, Chlamydia pneumoniae pneumoniae pneumonia and Legionella pneumophila pneumonia caused by the three pathogenic bacteria are atypical ...

AI Technical Summary

Problems solved by technology

[0009] 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

[0010] 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

[0011] In addition, the mixed infection of Mycoplasma pneumoniae, Chlamydia pneumoniae and Legionella pneumophila is increasing day by day, and the pneumonia caused by the three infections is not specific in terms of symptoms, signs and auxiliary examinations, and is easy to be misdiagnosed. The product cannot meet the needs of rapid detection and differentiation of three pathogens