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Method for quickly identifying food pathogenic bacteria subtype based on asymmetric polymerase chain reaction (PCR) combined test strip platform and kit

A technology for pathogenic bacteria and test strips, which is applied in the field of biological detection and can solve the problems of long inspection cycle and the need for instruments.

Active Publication Date: 2014-03-19
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have the disadvantages of long inspection period and expensive instruments.

Method used

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  • Method for quickly identifying food pathogenic bacteria subtype based on asymmetric polymerase chain reaction (PCR) combined test strip platform and kit
  • Method for quickly identifying food pathogenic bacteria subtype based on asymmetric polymerase chain reaction (PCR) combined test strip platform and kit
  • Method for quickly identifying food pathogenic bacteria subtype based on asymmetric polymerase chain reaction (PCR) combined test strip platform and kit

Examples

Experimental program
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Effect test

Embodiment 1

[0097] 1. DNA extraction of Listeria monocytogenes

[0098] The genomic DNA of Listeria monocytogenes (strain number CMCC54007, purchased from Guangzhou Institute of Microbiology) was extracted using TIANamp Bacteria DNA kit.

[0099] 2. Primer and probe design

[0100] Design two pairs of primers based on the PRS gene and ORF2819 gene fragments, and add a common sequence at the 5'end of each pair of primers. Design the probe on the test strip according to the amplified product. The base sequence and label of each sequence are shown in Table 1:

[0101] Table 1 The base sequences and labels of the probes on the amplified product design test strips are as follows

[0102]

[0103]

[0104] 3. Preparation of Nano Gold

[0105] Nano gold was prepared by citrate reduction method, and 100mL1mM HAuCl 4 Heat to a boil, and quickly add 10mL 38.8mM sodium citrate solution under rapid stirring. The color of the solution changes from golden yellow→gray→wine red→translucent red within 2 minutes. C...

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Abstract

The invention discloses a method for quickly identifying a food pathogenic bacteria subtype based on an asymmetric polymerase chain reaction (PCR) combined test strip platform and a kit, and belongs to the technical field of biological detection. The method comprises the main steps of: (1) extracting food pathogenic bacteria subtype DNA; (2) designing an amplification primer and a nucleic acid probe sequence; (3) performing amplification reaction based on asymmetric PCR; (4) preparing a nanogold probe; (5) preparing a colloidal gold nucleic acid test strip; (6) detecting a sample. According to the method, food pathogenic bacteria in food samples can be quickly, specially, sensitively, qualitatively and quantitatively detected and the special subtype of the pathogenic bacteria is determined; the probe is simply designed, the operation steps are simple and short, the product is a single chain, and the method is easy to detect and convenient to popularize.

Description

Technical field [0001] The invention relates to the technical field of biological detection, in particular to a method and a kit for quickly identifying subtypes of food pathogens based on asymmetric PCR combined with a test strip platform. Background technique [0002] Food-borne pathogens are the primary cause of food-borne diseases, which can cause great harm to human health and are a major hidden danger to food safety. In the past 20 years, food safety problems caused by food-borne pathogens have been increasing. Therefore, carrying out LM (Listeria monocytogenes) epidemiological research is of great significance for preventing the prevalence of food-borne listeriosis. Fast, highly specific, and highly sensitive detection of pathogenic subtypes of food pathogens can not only quickly diagnose the cause of disease, but also prevent intestinal infectious diseases and food poisoning in real life. [0003] At present, the detection methods of food-borne pathogenic bacteria subtype...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/686C12Q2531/107C12Q2563/137C12Q2565/625
Inventor 邢达周小明刘芳
Owner SOUTH CHINA NORMAL UNIVERSITY
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