RT-LAMP Primer Kit for Rapid PEDV Strain Differentiation

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Solution Overview

Problem

Current diagnostic methods for porcine epidemic diarrhea virus (PEDV) cannot differentiate between classical and variant strains, requiring time-consuming and labor-intensive gene sequencing, and there is a need for a rapid and efficient method to identify and distinguish between these strains.

Innovation Solution

A reverse transcription loop-mediated isothermal amplification (RT-LAMP)-based primer set and kit that includes specific primer pairs for simultaneous identification of PEDV classical and variant strains, utilizing a universal and identification LAMP reaction system for rapid detection and differentiation.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If traditional diagnostic methods (gene sequencing) are used to differentiate PEDV strains, then measurement precision is improved, but loss of time increases significantly

Engineering Contradiction:

Improvestrain differentiation accuracyVSAvoiddetection time

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The patent divides the detection process into two independent RT-LAMP reactions: one targeting the conserved M gene for general PEDV detection, and another targeting the variable S gene for strain differentiation. This segmentation allows simultaneous detection and differentiation without requiring sequential gene sequencing, thereby reducing detection time while maintaining accuracy.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent performs preliminary genetic analysis to identify specific sequence differences between classical and variant strains in the S gene. Based on this preliminary knowledge, specific primers are designed and prepared in advance, enabling direct differentiation through RT-LAMP without needing to perform time-consuming sequencing during actual diagnosis.

Inventive Principle:

Principle #10Preliminary action

2Measurement precision

If gene sequencing is performed for strain identification, then measurement precision is improved, but device complexity increases

Engineering Contradiction:

Improvestrain identification accuracyVSAvoidequipment requirements

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent replaces complex mechanical sequencing equipment with a biochemical RT-LAMP system. By using temperature-controlled amplification and fluorescent dyes instead of sequencing machines, the method achieves strain differentiation accuracy comparable to sequencing while eliminating the need for expensive, complex equipment.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the detection parameter from sequence reading (requiring sequencing equipment) to amplification efficiency and fluorescent signal intensity (detectable by simple thermocyclers or even visual inspection). This parameter change allows accurate strain identification using minimal equipment.

Inventive Principle:

Principle #35Parameter changes

3Productivity

If RT-LAMP with specific primers is used for strain differentiation, then productivity is improved, but manufacturing precision increases

Engineering Contradiction:

Improvedetection speedVSAvoidprimer design complexity

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent designs a universal primer set for the M gene that can detect all PEDV strains, combined with strain-specific primers for the S gene. This universal approach allows the same RT-LAMP platform to serve both general detection and specific differentiation functions, improving productivity without requiring entirely separate systems for each purpose.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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 RT-LAMP method provides rapid, accurate, and cost-effective detection of PEDV strains with high sensitivity and specificity, capable of differentiating between classical and variant strains in 40 minutes without complex equipment, reducing the risk of false positives and negatives.

Implementation Method 1

reverse transcription loop-mediated isothermal amplification (RT-LAMP)-based primer set, a kit, and a detection method for rapid and simultaneous identification of a classical strain and a variant strain of a PEDV

Methodology Applied

Scientific EffectLoop-mediated isothermal amplification:

Data Source

PatentUS20260103767A1Primer set and kit for simultaneous identification of classical strain and variant strain of porcine epidemic diarrhea virus (PEDV)

Publication Date: 2026.04.16 JIANGXI AGRICULTURAL UNIVERSITY

AI summary

A primer set and a kit is used for simultaneous identification of a classical strain and a variant strain of a porcine epidemic diarrhea virus (PEDV). The primer set includes primer set I and primer set II; where primer set I comprises the nucleotide sequences set forth in SEQ ID NO: 1 to SEQ ID NO: 4; and primer set II comprises the nucleotide sequences set forth in SEQ ID NO: 5 to SEQ ID NO: 8. This approach not only helps avoid false negatives or false positives in the detection of PEDV variant strain infections, but also enables the identification between classical strain and variant strain in PEDV. This is of significant importance for understanding the prevalence of PEDV and for the prevention and control of PEDV infections.