Mycoplasma pneumoniae detection device and detection method

A technology of Mycoplasma pneumoniae and a detection device, applied in the field of genetic engineering, can solve problems with high requirements, and achieve the effects of satisfying rapid detection, improving convenience, and improving generalizability

Pending Publication Date: 2022-06-03
NANHUA HOSPITAL AFFILIATED TO UNIV OF SOUTH CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

qPCR is the main method for early and rapid detection of Mycoplasma pneumoniae nucleic acid. It can perform relative and absolute quantification,

Method used

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  • Mycoplasma pneumoniae detection device and detection method
  • Mycoplasma pneumoniae detection device and detection method
  • Mycoplasma pneumoniae detection device and detection method

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experiment example

[0067] ERA / CRISPR-Cas12a double detection was performed on 62 suspected positive samples and 30 negative samples, and the sequences were extracted with QIAmpDNA Mini Kits. The above samples have been renumbered out of order to protect patient privacy. All tests were repeated three times, and the qPCR results were used as the gold standard to evaluate the coincidence rate (PPA / NPA) of the ERA / CRISPR-Cas12a dual system for clinical specimens.

[0068] The extracted DNA was subjected to real-time PCR with qPCR kit and amplified with ABI7500. The system is as follows: 38L of reaction solution, 2L of enzyme mixture, and 0.4L of internal standard substance are configured into enzyme mixture according to the reaction ratio. Add 10L of the sample to be tested into the amplification reaction tube, let it stand for 10 minutes, add 40L of enzyme mixture, and put it on the amplification instrument after spotting. The amplification program was: 50°C for 2 minutes; 94°C for 5 minutes; the...

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Abstract

The invention relates to the technical field of gene engineering, and provides a mycoplasma pneumoniae detection device and method.The mycoplasma pneumoniae detection device comprises a shell, a sample inlet and outlet module, a reaction container, a feeding module, a heating module, a fluorescence detection module and a control module; the amplification product and the LbCas12a-gRNA compound can be mixed and subjected to amplification circulation, and if the fluorescence effect judgment exists, the mycoplasma pneumoniae exists in the sample. According to the method, detection of the mycoplasma pneumoniae can be completed within a short time, and whether the mycoplasma pneumoniae sequence exists or not can be detected within a short time. The device is convenient to carry and store, and can meet the requirement of rapid detection.

Description

technical field [0001] The present invention relates to the technical field of genetic engineering, in particular, to a detection device and a detection method for Mycoplasma pneumoniae. Background technique [0002] Mycoplasma pneumoniae is the smallest prokaryotic microorganism, has no cell wall, is highly polymorphic, and is a common pathogen that causes atypical pneumonia and community-acquired pneumonia (CAP). The clinical symptoms are mainly cough, fever, sore throat and muscle pain, which are similar to other respiratory pathogens such as Staphylococcus aureus, Acinetobacter baumannii, influenza virus and SARS-CoV-2, and are easily misdiagnosed. [0003] The traditional identification of Mycoplasma pneumoniae includes isolation and culture, immunological detection and molecular biological detection. Molecular biology is the most important method for the detection of Mycoplasma pneumoniae, including polymerase chain reaction (PCR), real-time quantitative PCR (qPCR) an...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12M1/36C12M1/38C12Q1/689C12Q1/6844C12N15/11C12R1/35
CPCC12M41/36C12M41/12C12M41/00C12Q1/689C12Q1/6844C12Q2521/507C12Q2521/101C12Q2522/101C12Q2563/107
Inventor 何军邓仲良胡海洋胡茜盼苏晓玲
Owner NANHUA HOSPITAL AFFILIATED TO UNIV OF SOUTH CHINA
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