HPV Genotype Detection Using E1 Gene Primers and Probes
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Solution Overview
Problem
Current methods for detecting and typing genital human papillomavirus (HPV) genotypes are limited in sensitivity and specificity, particularly in detecting high-risk types associated with cancer, and require time- and cost-intensive sequencing and gel electrophoresis for typing.
Innovation Solution
The use of specific oligonucleotides as primers and probes that target the E1 gene region of HPV, allowing for the amplification and detection of a broad spectrum of HPV types, including high-risk genotypes, using PCR and nucleotide arrays for efficient typing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PCR amplification is used to detect HPV DNA, then sensitivity of detection is improved, but time and cost increase due to required sequencing and gel electrophoresis for typing
Solution Approach 1:
The invention divides the HPV detection process into two independent stages: (1) PCR amplification of the E1 gene region using universal primers to detect all HPV types, and (2) sequencing only the amplified E1 fragment for typing. This segmentation eliminates the need for time-consuming gel electrophoresis while maintaining high detection sensitivity and typing accuracy.
Solution Approach 2:
The invention extracts and sequences only the E1 gene region of HPV, which is highly conserved across all HPV types. By taking out just this specific region for sequencing instead of analyzing entire viral genomes or using gel electrophoresis, the method achieves accurate typing with significantly reduced time and cost while maintaining high detection sensitivity through PCR amplification.
2Measurement precision
If PCR amplification is used to detect HPV DNA, then detection sensitivity is improved, but cost increases due to sequencing requirements
Solution Approach 1:
The invention extracts and sequences only the E1 gene region of HPV, which is highly conserved across all HPV types. By taking out just this specific region for sequencing instead of analyzing entire viral genomes or using gel electrophoresis, the method achieves accurate typing with significantly reduced time and cost while maintaining high detection sensitivity through PCR amplification.
Solution Approach 2:
The invention changes the sequencing target from entire viral genomes or multiple gene regions to only the E1 gene region. This parameter change in the sequencing scope, combined with universal PCR primers targeting E1, maintains high detection sensitivity across all HPV types while dramatically reducing sequencing costs and time requirements.
3Adaptability or versatility
If universal primers targeting conserved regions are used, then broad spectrum detection is improved, but typing precision deteriorates due to sequence variability
Solution Approach 1:
The invention divides the HPV detection process into two independent stages: (1) PCR amplification of the E1 gene region using universal primers to detect all HPV types, and (2) sequencing only the amplified E1 fragment for typing. This segmentation eliminates the need for time-consuming gel electrophoresis while maintaining high detection sensitivity and typing accuracy.
Solution Approach 2:
The invention changes the sequencing target from entire viral genomes or multiple gene regions to only the E1 gene region, which is highly conserved across all HPV types. This parameter change ensures that universal primers can amplify all HPV types while the sequenced E1 region provides sufficient precision for accurate genotype identification.
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
Enables fast, reliable, and specific amplification and typing of HPV genotypes, including rare high-risk types, reducing the need for extensive sequencing and gel electrophoresis, and improving the detection of HPV infections relevant to cancer development.
Implementation Method 1
The use of specific oligonucleotides as primers and probes that target the E1 gene region of HPV, allowing for the amplification and detection of a broad spectrum of HPV types
Implementation Method 2
allowing for the amplification and detection of a broad spectrum of HPV types, including high-risk genotypes, using PCR
Implementation Method 3
using PCR and nucleotide arrays for efficient typing
Data Source
AI summary
The invention relates to oligonucleotides, which are suited as primers for amplifying DNA of genital human papilloma viruses (HPV), to oligonucleotides, which are suited for use as probes for typifying genital HPV genotypes, to methods for amplifying the DNA of genital human papilloma viruses (HPV), to methods for detecting and/or identifying genital HPV genotypes, to nucleotide microarrays containing the oligonucleotides, to kits and to the use of the oligonucleotides for amplifying or typifying genital HPV genotypes, for the diagnosis and/or early diagnosis of diseases and for producing agents for diagnosing diseases.