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LAMP method for detecting real-time turbidity of Listeria monocytogenes

A technology for mononucleosis and Listeria, applied in the field of microbial detection, can solve the problems of rapid detection of unfavorable foodborne diseases, low accuracy and sensitivity, and cumbersome operation

Inactive Publication Date: 2015-01-28
ZHOUSHAN INST OF CALIBRATION & TESTING FOR QUALITY & TECHNICAL SUPERVISION
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Problems solved by technology

[0003] At present, the detection of Listeria monocytogenes is still based on the traditional method, which not only has a long inspection cycle (4-7 days), cumbersome operation, but also has low accuracy and sensitivity, which is not conducive to the rapid detection of foodborne diseases

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  • LAMP method for detecting real-time turbidity of Listeria monocytogenes
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  • LAMP method for detecting real-time turbidity of Listeria monocytogenes

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Embodiment Construction

[0019] The method of the present invention will be described in detail below in conjunction with embodiments.

[0020] 1. Design of LAMP primer set for detecting Listeria monocytogenes

[0021] According to the hlyA gene of Listeria monocytogenes published in GeneBank, the present invention performs sequence conservation analysis, and screens out highly conserved sequences of Vibrio alginolyticus, using Primer Explorer V4.0 software (http: / / primer-explorer.jp / e / v4-manual / index.heml) for primer design. Designing suitable primers is the key to the LAMP reaction. The primers for the LAMP reaction of Vibrio alginolyticus are designed by considering factors such as base composition, GC content, secondary structure formation, and Tm value. The primers were synthesized by Bao Bioengineering (Dalian) Co., Ltd. The primers are shown in Table 1:

[0022] Table 1 LAMP primer sequence used to amplify the hlyA gene of Listeria monocytogenes

[0023]

[0024]

[0025] 2. Detection of the effect...

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Abstract

The invention aims at providing an LAMP method for rapidly detecting the real-time turbidity Listeria monocytogenes, for making up the defects of the prior art. The method comprises the following step: firstly, providing an LAMP primer group for detecting Listeria monocytogenes, wherein the nucleotide sequences of the LAMP primer group are as shown in SEQ ID NO:1-4. A set of LAMP primers are designed aiming at the hemolysin gene of the Listeria monocytogenes, a real-time turbidity instrument is adopted to detect the Listeria monocytogenes, and the specificity and the sensitivity of the method are verified, so that a rapid, specific and sensitive Listeria monocytogenes LAMP detection method is established.

Description

Technical field [0001] The invention belongs to the technical field of microbial detection, and specifically relates to a real-time turbidity LAMP rapid detection method of Listeria monocytogenes. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes) is a type of facultative cell-growing gram-positive bacteria with strong pathogenicity and can cause listeriosis in humans and animals. It is an important food-borne human and animal Composed of pathogenic bacteria. Listeria monocytogenes has low nutritional requirements and strong environmental adaptability. It can grow at 2 to 4°C or survive at -18°C, even under repeated freezing and thawing conditions. In recent years, the number of patients in European and American countries from eating foods contaminated by Listeria monocytogenes has been on the rise. The World Health Organization (WHO) has listed it as one of the four major pathogens in food since the 1990s, and has become food hygiene and safety in man...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2531/119
Inventor 黄朱梁薛超波王萍亚
Owner ZHOUSHAN INST OF CALIBRATION & TESTING FOR QUALITY & TECHNICAL SUPERVISION
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