Method for detecting pathogenic microorganisms by using PCR (polymerase chain reaction) enzyme-linked double-cross method

Abstract

The invention provides a method for detecting pathogenic microorganisms by using a PCR (polymerase chain reaction) enzyme-linked double-cross method, belongs to the technical field of clinical medical microorganism detection, and particularly relates to a synchronous qualitative and quantitative detection method for various microorganisms. In the method, DNA (deoxyribonucleic acid) amplification, DNA hybridization and enzyme-linked immunosorbent assay reaction are used, a solid-phase surface enveloping technology is adopted, a specificity conservative segment is amplified, an amplified product is combined to a capturing probe and is adsorbed to a solid-phase surface, far-end hybridization of an enzyme-linked probe is carried out, and a quantitative color product is generated by specific antibody recognition and reaction of enzyme and a substrate. The detected pathogenic microorganisms comprise EB (Epstein Barr) viruses, giant cell and cell viruses, herpes simplex viruses 1 and herpes simplex viruses 2. Various microorganisms of a clinical sample are detected simultaneously, and four types of viruses are screened by a reaction. The method has the advantages that the time and the cost are saved, application is flexible, the repeatability, the stability, the sensitivity and the specificity are high, and the method is simple and is easy to implement. Moreover, the method is suitable for widely detecting pathogenic microorganisms and is particularly suitable for a basic medical institution.

Description

technical field [0001] The invention relates to the field of microbial detection in clinical medicine, water, environment and the like, in particular to the field of quantitative and qualitative detection of microorganisms. Background technique [0002] At present, domestic and foreign detection methods for clinical microbiology mainly include some simple molecular biology and immunological methods such as traditional biochemical identification method, immunological detection technology, nucleic acid molecular hybridization technology, PCR technology, and gene chip technology. [0003] The traditional biochemical identification method needs to inoculate the sample to be tested on the medium for isolation and culture, and to identify the pathogen species through a series of tedious characteristics and results of physiological and biochemical experiments. Although this method is a commonly used and classic method in microbiology laboratories identification method, but the proc...

AI Technical Summary

Problems solved by technology

Its sequencing principle is the same as that of nucleic acid hybridization, but it solves the shortcomings of traditional nucleic acid hybridization technology such as complicated operation, low detection efficiency and low degree of automation

The gene chip technology is expensive, and there are still some problems to be solved, such as improving the specificity of the chip and the sensitivity of the detection signal, reducing the production cost of the chip, etc., and most chips require expensive detection instruments. It is mainly limited to laboratory research and has not been widely used in the detection and identification of clinical pathogenic microorganisms

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