Multiplex PCR Kit for Listeria Serotype 4H Detection

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Solution Overview

Problem

Current methods for detecting Listeria monocytogenes, particularly serotype 4h, are time-consuming, labor-intensive, and unable to quickly distinguish hyper-virulent strains, posing challenges for rapid and effective food safety monitoring.

Innovation Solution

A multiplex PCR detection kit utilizing specific gene targets Imo1210 and xysn_1693 with corresponding primers for rapid detection of Listeria monocytogenes serotype 4h, employing PCR technology to amplify and identify the genes, thereby determining the presence of the serotype.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional biochemical identification experiment is used after enrichment culture and selective culture, then detection accuracy is improved, but detection time and labor consumption increase significantly

Engineering Contradiction:

Improvedetection accuracyVSAvoiddetection time

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The patent replaces conventional biochemical identification methods with molecular biology-based PCR detection. Specifically, it uses multiplex PCR amplification targeting serotype-specific genes (such as inlA, plcB, and other virulence factor genes) to directly identify Listeria monocytogenes serotypes from enriched samples, eliminating the need for time-consuming biochemical tests while maintaining or improving detection accuracy.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent performs serotype-specific PCR amplification during or immediately after the enrichment culture phase. By designing primers that target serotype-specific genetic markers, the method can preliminarily identify the serotype of Listeria monocytogenes before final confirmation, significantly reducing the overall detection time while preserving accuracy.

Inventive Principle:

Principle #10Preliminary action

2Measurement precision

If conventional biochemical identification method is used, then detection accuracy is improved, but operational complexity and labor intensity increase

Engineering Contradiction:

Improvedetection accuracyVSAvoidoperational complexity

Core Design Contradiction:

Measurement precisionVSEase of operation

Solution Approach 1:

The patent replaces complex biochemical identification procedures with a streamlined molecular detection system. The multiplex PCR assay uses specifically designed primer sets that amplify serotype-specific genetic sequences, allowing straightforward identification through gel electrophoresis or real-time fluorescence detection, thereby reducing operational complexity while maintaining high accuracy.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent develops a universal detection platform that can identify multiple Listeria monocytogenes serotypes simultaneously using a single multiplex PCR reaction. This multi-functional system uses a combination of primers targeting different serotype-specific genes, enabling one assay to perform the work of multiple separate biochemical tests, thus simplifying operations while preserving detection accuracy.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Measurement precision

If conventional detection method is used, then comprehensive identification is achieved, but ability to quickly distinguish hyper-virulent strains is lost

Engineering Contradiction:

Improveidentification comprehensivenessVSAvoiddetection speed

Core Design Contradiction:

Measurement precisionVSSpeed

Solution Approach 1:

The patent focuses on detecting specific local genetic characteristics associated with hyper-virulence in Listeria monocytogenes. By designing primers that target virulence factor genes (such as inlA, inlB, plcB, hly, and other pathogenicity island genes), the method can specifically identify hyper-virulent strains among general Listeria populations, enabling rapid differentiation based on these critical local genetic features while maintaining comprehensive identification capabilities.

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent performs targeted amplification of virulence factor genes during the early enrichment phase. By using PCR to detect the presence of specific virulence-associated genetic markers, the system can preliminarily distinguish hyper-virulent strains from non-hyper-virulent ones before final confirmation, significantly accelerating the detection process while preserving comprehensive identification accuracy.

Inventive Principle:

Principle #10Preliminary action

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The kit provides a rapid, accurate, and specific method for detecting Listeria monocytogenes serotype 4h, with no false positives and high sensitivity, significantly reducing detection time and operational complexity compared to traditional methods.

Implementation Method 1

detects the gene Imo1210 and the gene xysn_1693 using PCR technology

Methodology Applied

Scientific EffectPCR amplification:

Data Source

PatentUS11827945B2Multiplex PCR detection kit for listeria monocytogenes serotype 4H

Publication Date: 2023.11.28 YANGZHOU UNIV

AI summary

The present invention provides a multiplex PCR detection kit of Listeria monocytogenes serotype 4h. The kit includes a gene Imo1210 detection primer and a gene xysn_1693 detection primer. The present invention establishes a multiplex PCR method for rapidly detecting Listeria monocytogenes serotype 4h by using two pairs of primers.