Method and kit for fast detection of human chlamydia pneumoniae antigen based on magnetic resolution and quantum dot labelling

Abstract

The invention provides a method for detection of human chlamydia pneumoniae antigen based on magnetic resolution and quantum dot labelling. The method comprises the following steps: (1) preparing anti-human chlamydia pneumoniae immune nano magnetic beads; (2) preparing quantum dot labelled anti-human chlamydia pneumoniae nanoprobes; (3) dissolving a sample to be detected with a sample treating solution, adding the anti-human chlamydia pneumoniae immune nano magnetic beads into the dissolved solution, carrying out magnetic separation after sufficient mixing and reaction, washing with a PBST buffer solution, adding the quantum dot labelled anti-human chlamydia pneumoniae nanoprobes into the obtained precipitate, carrying out magnetic separation after a reaction, washing with the PBST buffer solution, and detecting the fluorescence value with a fluorescence micro-plate reader. The method is accurate, fast, and high in sensitivity, and has very high practical value in the aspects of clinical diagnosis, etiological differentiation, epidemiological surveys and the like of human chlamydia pneumoniae.

Description

Technical field [0001] The invention relates to the technical field of medical detection, in particular to a rapid detection method and a detection kit for detecting human Chlamydia pneumoniae (Cpn) antigen based on magnetic separation and quantum dot labeling, as well as preparation and use methods of the detection kit. Background technique [0002] Chlamydiapneumoniae (Cpn), as an important respiratory pathogenic microorganism of the genus Chlamydia, has been officially named by Grayston et al. in the mid-1980s and has been extensively studied by researchers in the following period. It is only known that humans are the host of the pathogen, and the mode of infection may be spread from human to human through respiratory secretions. Children under 5 years of age are rarely infected. Children over 8 years of age and young people are susceptible to infection, especially in crowded areas such as homes, schools, and barracks. Seroepidemiological investigations have confirmed that at ...

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Problems solved by technology

However, the PCR experiment has special requirements for the laboratory, the sample processing, amplification and detection requirements are strict, and it is prone to false positives, so it cannot be used as a commonly used clinical diagnostic method in my country.

[0005] Antigen detection methods include separation and culture of chicken embryo yolk sac and cell culture method (two kinds of cells, HL and Hep-2, are commonly used), and then use fluorescent-labeled or enzyme-labeled antibodies to detect Chlamydia in the specimen, but this method has complicated operation steps and cell culture Obvious defects such as long time, not suitable for clinical application

In recent years, it has also been tried to be directly applied to the detection of clinical specimens, and its sensitivity is related to specimen collection: EIA can be used to detect the presence of Chlamydia pneumoniae antigen after smears of sputum and throat swabs, but its sensitivity, specificity and reproducibility are not ideal , and is limited to acute respiratory infection; enzyme-linked immunosorbent assay (ELISA) can also directly detect pathogens, mainly detecting Chlamydia pneumoniae outer membrane protein-2, the antibody used is an anti-Chlamydia specific antibody, which cannot directly recognize Chlamydia pneumoniae, and Sensitive but less specific than the "gold standard" (MIF)

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