Kit for instantly detecting vaginitis pathogen gene

Abstract

The invention discloses a kit for instantly detecting a vaginitis pathogen gene. The kit is used for amplification and detection of a gene at constant temperature. The kit can perform rapid fluorescence detection through matching between an isothermal amplification reaction reagent and a two-color fluorescent labeled probe. The isothermal amplification system of the kit really achieves gene amplification reaction at the same temperature, does not depend on ATP, creatine phosphate and other high-energy energy molecules, is simpler in composition, lower in cost and more convenient to use compared with an existing isothermal amplification system. The isothermal amplification system is suitable for amplification of complex templates, is high and stable in amplification speed, can complete amplification reaction within 15 minutes, high in amplification sensitivity and accuracy, does not produce mismatch and is good in amplification effect.

Description

technical field [0001] The invention relates to a vaginitis pathogen gene instant detection kit, in particular to a constant temperature amplification technology independent of high-energy energy molecules. Background technique [0002] Vaginitis is the most common disease in obstetrics and gynecology patients. About 5 to 10 million women suffer from this disease every year, and 75% of women suffer from it at least once in their lifetime. Traditional vaginitis diagnostic methods mainly rely on patient complaints, vaginal examination, and laboratory tests. The symptoms of vaginitis lack specificity, so in the absence of laboratory confirmation, it is difficult to make a diagnosis based on the patient's complaint and other examinations. Even if most patients with vaginitis have repeated infections, the cause of the disease still cannot be confirmed, and most patients with vaginitis are mixed infections, so the diagnosis of vaginitis is very difficult. The most common types o...

AI Technical Summary

Problems solved by technology

[0004] The detection of vaginitis-causing pathogen DNA often uses traditional PCR technology. The detection program needs to set up multiple cycles, and each cycle includes denaturation, annealing, and extension of nucleic acids, which leads to several limitations in the technical application of PCR: First, PCR The technology requires expensive laboratory instruments. At the same time, the instrument must have a fine temperature control program and a heated template, so as to realize the denaturation of the DNA template strand at a very high temperature, and the primer probe anneals at a lower temperature to extend the template, so that the temperature can be repeated. After changing dozens of cycles, the amount of template can be enlarged

Since each cycle time is short, the instrument must be able to quickly and accurately raise and lower the temperature, which requires a silver or gold heating module, which increases the cost of instrument development

Second, the polymerase in the PCR amplification system must have the ability to withstand high temperature, otherwise new enzymes must be added in each cycle to achieve the amplification of the next cycle, which is very easy to cause pollution and increase the cost

Excess primers can mismatch with the template, misprime amplification, primer dimers, etc., which in turn inhibit PCR amplification, especially when the amount of template is low, which can exacerbate this situation

Images