Primer, fluorescence probe and kit for quantitative detection of streptococcus pneumonia nucleic acid and detection method of streptococcus pneumonia nucleic acid

Abstract

The invention provides a primer, a fluorescence probe and a kit for the quantitative detection of streptococcus pneumonia nucleic acid and a detection method of streptococcus pneumonia nucleic acid, wherein the primer comprises a forward primer and a backward primer; and the kit is used for the quantitative detection of streptococcus pneumonia nucleic acid and comprises the primer and the fluorescence probe, and PCR reaction liquid, a DNA extraction solution, a negative quality control material, a positive quality control material, a critical positive quality control material and a working standard. The method for the quantitative detection of streptococcus pneumonia nucleic acid through using the primer, the fluorescence probe and the kid comprises the following steps: step 1, sample collection; step 2, sample processing; step 3, sample application; step 4, PCR amplification; and step 5, analysis and judgment. According to the real-time TaqMan fluorescence quantitative PCR provided by the embodiment of the invention, the primer and the fluorescence probe have high specificity and high sensitivity, the kit has precise quantification, and the detection method can rapidly detect the streptococcus pneumonia.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a primer for quantitative detection of Streptococcus pneumoniae nucleic acid, a fluorescent probe, a kit and a detection method thereof. Background technique [0002] Streptococcus pneumoniae (SP) belongs to Streptococcus family Streptococcus bacteria, Gram stain positive. It does not produce endotoxins and exotoxins, and its pathogenicity is mainly due to the invasion effect of the capsule: the non-capsulated variant strain is non-virulent, and the capsulated pneumococcus can resist the phagocytosis of phagocytes, which is conducive to the colonization of bacteria in the host and reproduce. Streptococcus pneumoniae can be divided into more than 90 serotypes according to the polysaccharide antigenicity of the bacterial capsule, and different serotypes have different pathogenicity. [0003] 75% of adult Streptococcus pneumoniae pneumonia and more than 50% of severe ...

AI Technical Summary

Problems solved by technology

[0004] However, there are still deficiencies in the above methods: using traditional culture methods (national standard method), latex agglutination method, immunochromatography and nucleic acid amplification technology detection, there will be false positive or false negative results, which will affect the accuracy of the experimental results Automated method Because the automatic identification system identifies bacteria according to the background information provided in the database, the incompleteness of the database information will directly affect the accuracy of identification, and there is currently no identification system that can include all detailed identification information; nucleic acid amplification Although the results of the amplification technology are better than the above four methods, the PCR amplification products need to be post-processed, and the contamination of the PCR products during the post-processing will still lead to false positive results, and will pollute the environment and have potential risks to the experimenters. Harm: Although real-time fluorescent quantitative PCR technology solves the problem of nucleic acid amplification technology pollution, and the detection results are more specific than nucleic acid amplification technology, it still cannot achieve 100% specificity

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