Method and chip for detecting bacterial nucleic acid and application of chip in decayed tooth detection

Abstract

The invention provides a method and a chip for detecting bacterial nucleic acid and application in decayed tooth detection. The method specifically comprises the following steps: S1, centrifugally mixing a probe, an amplification reagent, to-be-detected bacteria and a developing reagent uniformly, wherein wthe probe is a hairpin probe with a stem-loop structure and is composed of three fragments: (1) a G-quadruplex sequence is designed and formed at the 3' tail end, (2) nine basic groups at the 5' tail end can be complementarily paired with part of the G-quadruplex sequence to hinder the formation of the G-quadruplex, and (3) an annular region; S2, carrying out an amplification reaction; and S3, carrying out quantitative detection on the 16S rRNA through a colorimetric reaction. According to the invention, a centrifugal micro-fluidic chip is combined with a colorimetric method based on double-strand specific nuclease-assisted target circulation and probe cutting, and is used for sensitively detecting and distinguishing streptococcus mutans and streptococcus sobrinus in decayed teeth.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis, and in particular relates to a method for detecting bacterial nucleic acid, a chip and an application in dental caries detection. Background technique [0002] Dental caries is a chronic infectious disease that occurs in teeth, and is considered to be one of the most common infectious diseases in humans. The onset trend is progressive and irreversible, directly leading to the loss of tooth tissue or even the loss of the entire tooth, and a series of infections and pains. Streptococcus mutans and Streptococcus farnus have been recognized as pathogens of dental caries in humans. Some studies have shown that the prevalence of Streptococcus aureus is more correlated with caries activity than that of Streptococcus mutans, which also has a higher acid production capacity compared with Streptococcus mutans. According to reports, the incidence of dental caries in children carrying Streptococcus mutan...

AI Technical Summary

Problems solved by technology

Later, polymerase chain reaction (PCR) was used to detect and distinguish Streptococcus mutans and Streptococcus farus. However, PCR requires sophisticated instruments to precisely control the reaction temperature, which is inconvenient to carry and expensive, and is not suitable for clinical dental treatment. diagnosis

Subsequently, loop-mediated isothermal amplification (LAMP) was developed, which greatly improved the sensitivity of detection, however, these methods involved complex primer / template design and time-consuming experiments with inherently nonspecific background amplification program

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