Method and system for detecting brucella based on micro-fluidic chip-PCR (Polymerase Chain Reaction) technology

A technology of Brucella and detection system, applied in a field of detection of Brucella, can solve the problems such as no application report of the detection method of Brucella, and achieve the effect of safety and good accuracy of the detection method

Pending Publication Date: 2022-05-13
杜彦丹 +1
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Problems solved by technology

[0005] However, there is no application report of Brucella detection method based on MC-PCR technology at present

Method used

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  • Method and system for detecting brucella based on micro-fluidic chip-PCR (Polymerase Chain Reaction) technology
  • Method and system for detecting brucella based on micro-fluidic chip-PCR (Polymerase Chain Reaction) technology
  • Method and system for detecting brucella based on micro-fluidic chip-PCR (Polymerase Chain Reaction) technology

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Embodiment 1

[0052] A kind of brucella detection technology based on MC-PCR technology is provided in the present embodiment, wherein be Beijing Bohui innovation brucellosis detection microfluidic nucleic acid chip and Beijing Bohui innovation full-automatic nucleic acid chip detector (model BHF -VI) Supporting use for detecting Brucella.

[0053] In this embodiment, the reagents, consumables and main components used are shown in Table 1.

[0054] Table 1

[0055]

[0056]

[0057] Remarks: ①The packaging volume deviation of the above liquid reagent components shall not exceed ±10% of the nominal specification;

[0058] ② Different batches of Brucella nucleic acid detection kits shall not be mixed;

[0059] ③ The lysate and chromogenic solution should be stored away from light.

[0060] ④External standard quality control products include positive control and negative control, which need to be processed during each experiment, and the method is the same as the sample operation.

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Abstract

The invention provides a method and a system for detecting brucella based on a micro-fluidic chip-PCR (Polymerase Chain Reaction) technology. The method for detecting Brucella comprises the following steps: performing target DNA extraction, PCR amplification and reverse dot blot hybridization on a to-be-detected sample, wherein the target DNA extraction is performed on the to-be-detected sample by adopting a NaI magnetic bead method; carrying out PCR (Polymerase Chain Reaction) amplification by taking the extracted DNA as a template and adopting a primer designed aiming at the Brucella BCSP-31 gene; carrying out reverse dot blot hybridization on a PCR amplification product on a hybridization membrane; wherein a detection probe designed aiming at a Brucella BCSP-31 gene is fixed on the hybridization membrane. The detection method disclosed by the invention can be fully automatically carried out by utilizing the micro-fluidic chip, is safe, rapid, timely and accurate, and the application of the technology disclosed by the invention can improve the diagnostic rate and diagnostic accuracy of Brucella, and plays an important role in preventing and treating Brucella diseases.

Description

technical field [0001] The present invention relates to a method and system for detecting Brucella, in particular to a method and system for detecting Brucella based on microfluidic chip-PCR technology. Background technique [0002] Brucella (Brucella, also known as Brucella) is a Gram-negative non-motile bacterium, which invades the body and causes infection-allergic zoonotic infectious diseases——Brucella sick. Brucellosis is clinically characterized by long-term fever, hyperhidrosis, fatigue, joint pain, hepatosplenomegaly, and lymph node enlargement, and the crowd is susceptible. It is very important to detect and diagnose the pathogen early, quickly and accurately. The detection of Brucella is the gold standard for diagnosing brucellosis. However, due to the high nutritional requirements and slow growth of Brucella culture, the traditional culture detection method often delays diagnosis and early treatment, so it cannot be used as a diagnostic tool. The main method of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6806C12Q1/686C12Q1/6837C12N15/11C12R1/01
CPCC12Q1/689C12Q1/6806C12Q1/686C12Q1/6837C12Q2523/308C12Q2521/537C12Q2527/125C12Q2565/629C12Q2545/101C12Q2545/113
Inventor 杜彦丹赵志军李翔董占柱孙欣陈昊包春喜郝明媛李英智牛艺卿
Owner 杜彦丹
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