Pathogenic microorganism nucleic acid amplification-free detection and typing method

Abstract

The present invention discloses a pathogenic microorganism nucleic acid amplification-free detection and typing method, and related kits thereof, wherein a plurality of probes and a fluorescence quantum dot layer-by-layer assembly technology are combined to achieve pathogenic microorganism nucleic acid amplification-free detection and typing. According to the method, low concentration nucleic acid can be directly detected without amplification; with the plurality of the probes, the easily-appearing false positive problem during a signal amplification process is overcome so as to improve detection accuracy; and with the technology, real time pathogenic microorganism copy number detection and synchronization genotyping can be achieved, speed is fast, and cost is low.

Description

technical field [0001] The invention belongs to the field of medical biology, and in particular relates to a direct detection and typing method and kit of pathogenic microorganism nucleic acid without amplification. Background technique [0002] Infectious diseases are one of the most important diseases that seriously endanger human health. According to the statistics of the National Center for Disease Control and Prevention (CDC): In 2011, there were 6.32 million cases of legal infectious diseases in my country and 15,000 deaths. Among them, the incidence of viral hepatitis, tuberculosis and syphilis ranks the top three, accounting for 85.41% of the total incidence of Class B infectious diseases. And the incidence of blood-borne infectious diseases such as hepatitis B and hepatitis C is increasing year by year. The above data show that viral hepatitis B still occupies the first place in the incidence of infectious diseases in my country, and its number of cases also accou...

AI Technical Summary

Problems solved by technology

These PCR-based amplification techniques for HBV detection and typing still have the following deficiencies: (1) the amplification conditions are extremely strict, and false positives or false negatives are easily produced

(2) Synchronous amplification of multiple genotypes often leads to competitive inhibition of high-concentration templates against low-concentration templates, resulting in false negative results for low-concentration samples

(3) Barriers to the core intellectual property rights of PCR-related technologies lead to high prices for related reagents and equipment, increasing medical costs and patient burdens

However, in experiments, we found that traditional fluorescent dyes are easily bleached, making it difficult to detect clinical samples

[0006] However, the sequence homology between the A-H subtypes of HBV is very high, and the use of nucleic acid hybridization technology to type them requires the preparation of highly specific probes

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