Method for detecting mycoplasma pneumoniae antibody, kit for detection and preparation method of kit

Abstract

The invention relates to the technical field of biology, in particular to a method for detecting a mycoplasma pneumoniae antibody, a kit for detection by adopting the method and a preparation method of the kit. The method comprises the steps as follows: fixing anti-human IgM (immunoglobulin m) and anti-human IgG (immunoglobulin g) to a membrane respectively; adding a serum to be detected; after the mycoplasma pneumoniae antibody in the serum to be detected is bonded with the anti-human IgM and the anti-human IgG fixed to the membrane, adding an enzyme-labeled mycoplasma pneumoniae antigen solution; and adding a color developing solution for developing color and displaying a result at last. According to the method, the process of detecting the mycoplasma pneumoniae antibody by an enzyme-labeled mycoplasma pneumoniae antigen is quick and efficient, the quality guarantee period of the kit is prolonged, the detection sensitivity is improved, and the cost of the kit is reduced.

Description

Technical field [0001] The present invention relates to the field of biological technology, in particular to a method for detecting Mycoplasma pneumoniae antibodies, a kit for detection using this method, and a method for preparing the kit. Background technique [0002] Mycoplasma pneumoniae (Mycoplasma pneumoniae, Mp) is the pathogen of human primary atypical pneumonia. It can also cause other respiratory tract infectious diseases and even other tissue diseases of the body, and its incidence is increasing and there is a trend of epidemic. Because Mp has no cell wall, the treatment of infection caused by Mp is different from other bacterial and viral infections. Therefore, the pathogenic diagnosis of Mp infection is of great significance for the timely and correct treatment of the disease. [0003] Although there are many methods for detecting Mycoplasma pneumoniae, there is a lack of more rapid, simple and reliable methods. X-ray signs lack specificity, and must be differentiated...

AI Technical Summary

Problems solved by technology

[0004] The specificity and sensitivity of CAT are the lowest, so its clinical diagnostic value is not great; although the gold standard spot method is simple and quick to operate, its sensitivity is slightly low, and there is a possibility of missed diagnosis; the preparation and operation of gelatin particle agglutination test reagents are cumbersome and time-consuming , and requires professional operators

Enzyme-linked immunosorbent assay (ELISA) has the advantages of specificity, sensitivity and simplicity, and has been used to check various antibodies or antigens. However, this method has the following problems: (1) Each batch of tests starts from adding samples, incubating and washing plates. There are many steps, the operation is cumbersome, the time is long, and special operators and special instruments are needed, so it is difficult to carry out in primary medical units.

(2) The chromogenic components A and B need to be added separately during the detection, which increases the labor intensity and the cost of the kit to a certain extent

Patent application number 200710075306.3 discloses a color development system that can make chromogen and peroxide coexist within a certain period of time, but the key component in this system—chromogen protective agent sodium thiosulfate, invented here An acidic environment with a pH lower than 6.0 is not conducive to long-term stability. If the solution system has CO2, microorganisms or long-term exposure to light, it is not conducive to the stability of sodium thiosulfate, so it cannot play a role in chromogenic substances such as TMB. Effective Antioxidant Protection

Among them, Bassoon-1789, which is used to absorb ultraviolet rays, can protect against ultraviolet rays, but it turns red when it encounters metal ions, which interferes with the test results. Therefore, additional requirements for reagent purity and operation are added during the preparation process.

(3) The ELISA method cannot achieve rapid simultaneous detection of two antibodies

[0005] At present, the sodium periodate method is mostly used in HRP-labeled antibody technology. The technical process of a few scholars using HRP to label antigens is basically similar to the process of labeling antibodies. However, the following problems exist in this labeling process: (1) pH needs to be adjusted repeatedly during labeling, and The pH value adjustment range is wide, which inevitably affects the enzyme activity; (2) the labeling process is cumbersome and time-consuming; (3) the cross-linking of the enzyme itself is inevitable during the labeling process; (4) the label is not stable enough to store, Repeated freezing and thawing will reduce the activity of the marker

The storage buffer of some diagnostic kits does not give specific components for commercial reasons, which brings inconvenience to the development and application of the kits

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