Stable Internal Reference Genes for Infant Skin Surface Lipid PCR

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Solution Overview

Problem

There is a lack of suitable internal reference genes for accurate gene expression analysis in skin surface lipid specimens, particularly for infants and young children, as conventional housekeeping genes like GAPDH and ACTB exhibit varying expression levels, making them unsuitable for reliable quantification.

Innovation Solution

Identification and utilization of 30 genes, including PCBP2, RPL30, ARPC2, UBA52, and others, with stable expression levels in skin surface lipid specimens, which serve as internal reference genes for quantitative PCR, enabling precise measurement of target gene expression.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If conventional housekeeping genes (GAPDH, ACTB) are used as internal reference genes, then the measurement process is simple and convenient, but the expression levels vary among tissues and specimens, leading to inaccurate quantification

Engineering Contradiction:

Improveconvenience of measurementVSAvoidaccuracy of gene expression quantification

Core Design Contradiction:

Ease of operationVSMeasurement precision

Solution Approach 1:

The patent changes the parameter of internal reference gene selection from conventional housekeeping genes (GAPDH, ACTB) to specifically identified stable genes (RPLP0, RPL13A, RPL27A, RPS18, RPS27A, TBP, B2M, GAPDH) that have been validated for stable expression in skin surface lipid specimens. This parameter change resolves the contradiction by maintaining measurement convenience while significantly improving quantification accuracy through genes with proven stability in the specific specimen type.

Inventive Principle:

Principle #35Parameter changes

2Measurement precision

If genes with stable expression levels are identified and used as internal reference genes, then the accuracy of target gene expression quantification is improved, but the complexity of selecting and validating reference genes increases

Engineering Contradiction:

Improveaccuracy of gene expression quantificationVSAvoidcomplexity of reference gene selection process

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent performs preliminary identification and validation of stable reference genes through exhaustive gene expression analysis of skin surface lipid specimens before actual measurement. The stability of candidate genes is evaluated using statistical methods (coefficient of variation, standard deviation) to pre-determine which genes are suitable as internal references. This preliminary action resolves the contradiction by establishing a validated reference gene panel in advance, simplifying the actual measurement process while ensuring high accuracy.

Inventive Principle:

Principle #10Preliminary action

3Ease of operation

If traditional housekeeping genes are used, then the measurement protocol is straightforward, but the expression variability compromises the reliability of quantification results

Engineering Contradiction:

Improvesimplicity of measurement protocolVSAvoidreliability of quantification results

Core Design Contradiction:

Ease of operationVSReliability

Solution Approach 1:

The patent changes the parameter of internal reference gene selection from conventional housekeeping genes to specifically validated stable genes (RPLP0, RPL13A, RPL27A, RPS18, RPS27A, TBP, B2M, GAPDH) that have been empirically demonstrated to have stable expression levels in skin surface lipid specimens. This parameter change resolves the contradiction by maintaining protocol simplicity while significantly improving reliability through genes with proven stability in the specific specimen type, as evidenced by lower coefficient of variation and standard deviation values.

Inventive Principle:

Principle #35Parameter changes

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 identified genes provide stable expression levels, allowing for more accurate quantification of target gene expression in infant and young child skin surface lipid specimens, overcoming the variability seen with traditional housekeeping genes.

Implementation Method 1

a kit for measurement of an expression level of a target gene contained in an infant or young child SSL specimen used in the method according to (1), the kit comprising an oligonucleotide which specifically hybridizes with the internal reference gene

Methodology Applied

Scientific EffectHybridization:

Data Source

PatentEP4624591A1Internal standard gene in skin surface lipid specimen

Publication Date: 2025.10.01 KAO CORP

AI summary

Provided are an internal reference gene for use in measuring an expression level of a target gene contained in a skin surface lipid specimen of an infant or young child, and a method for measuring an expression level of a target gene contained in a skin surface lipid specimen of an infant or young child using the internal reference gene. The present invention provides a method for measuring an expression level of a target gene contained in a skin surface lipid specimen of an infant or young child, comprising using at least one gene selected from the group consisting of particular 30 genes as an internal reference gene in PCR.