Multi-species Reference Ladder and Multiplex PCR Kit for Forensic Species Identification
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Solution Overview
Problem
Current forensic techniques lack a streamlined process for rapid species identification in animal-related crime scenes, as existing methods are laborious and often fail to discriminate between domestic species due to limited availability of non-human species-specific kits and challenges with sample mixtures.
Innovation Solution
Development of a multi-species reference ladder and multiplex PCR kit that allows for simultaneous species identification using mitochondrial DNA cytochrome b gene amplicons, separable by capillary or gel electrophoresis, enabling the differentiation of multiple species in a single test.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple species-specific kits are used for species identification, then species discrimination capability is improved, but device complexity and analysis time increase
Solution Approach 1:
The patent combines multiple species-specific identification capabilities into a single multi-species reference ladder and multiplex PCR kit. The reference ladder contains DNA fragments from multiple species (human, dog, cat, cow, pig, sheep, horse, chicken, turkey, rabbit, guinea pig) with distinct size ranges, allowing simultaneous identification of multiple species in one test rather than requiring separate kits for each species.
Solution Approach 2:
The multiplex PCR kit is designed with universal primers that can amplify DNA from all target species simultaneously, and the reference ladder provides a universal size standard for comparing amplicons across all species. This multi-functional system replaces the need for multiple single-function species-specific kits.
2Reliability
If traditional sequential testing methods are used, then testing thoroughness is improved, but productivity and analysis time decrease
Solution Approach 1:
The patent merges multiple sequential testing steps into a single multiplex PCR reaction. By running all species-specific amplifications simultaneously in one reaction tube with optimized primer concentrations, the system maintains thorough species identification while reducing analysis time from multiple sequential tests to one parallel test.
Solution Approach 2:
The reference ladder is prepared in advance with DNA fragments of known sizes from all target species, creating a comprehensive size standard that allows direct comparison of all sample amplicons in a single electrophoresis run, eliminating the need for repeated testing and comparison steps.
3Measurement precision
If sample consumption is increased for thorough analysis, then measurement accuracy is improved, but loss of substance increases
Solution Approach 1:
The multiplex PCR kit uses optimized primer concentrations where the universal primers are present in excess relative to species-specific primers, allowing efficient amplification of target DNA from limited samples. This partial action approach ensures sufficient amplification product for accurate identification while minimizing the amount of starting DNA required.
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
This approach provides a quick and robust method for identifying 11 domestic animal species and human, reducing the need for repeated testing and minimizing reagents and analysis time, thereby enhancing the efficiency of species identification in forensic investigations.
Implementation Method 1
multiplex PCR kit that allows for simultaneous species identification using mitochondrial DNA cytochrome b gene amplicons
Implementation Method 2
separable by capillary or gel electrophoresis
Implementation Method 3
separable by capillary or gel electrophoresis
Data Source
AI summary
The subject invention provides multi-species reference ladders, multiplex PCR kits, and methods for multi-species identification using the multi-species reference ladders and multiplex PCR kits. Advantageously, the multi-species reference ladder and multiplex kit allows for a stream-lined analysis process and rapid species identification in a single test.