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Rapid genotyping of 32 insertion/deletion panel for human identification using fluorogenic probes-based multiplex
Yong-Jun Han1, Li-Yan Liu1, Zhen Rong1
1Bioinformatics Center of AMMS, Beijing 100850, China.
Analytical Biochemistry
|June 14, 2023
Summary
A new method for insertion and deletion (InDel) genotyping offers rapid, accurate, and cost-effective DNA analysis. This portable approach enhances forensic genetics applications for personal identification and paternity testing.
Area of Science:
- Forensic Genetics
- Molecular Biology
- Biotechnology
Background:
- Insertion and deletion (InDel) polymorphisms are valuable in forensic genetics due to low mutation rates and small amplicons.
- Current capillary electrophoresis and next-generation sequencing methods for InDel detection are time-consuming, costly, and complex.
- A need exists for a reliable, rapid, sensitive, and economical genotyping method for InDels.
Purpose of the Study:
- To develop a rapid, cost-effective, and portable method for InDel genotyping.
- To establish a validated InDel panel for forensic applications.
Main Methods:
- Development of a 32 InDel panel using fluorogenic probes-based multiplex real-time PCR.
- Utilized a microfluidic test cartridge and a portable real-time PCR instrument.
- Validation studies included concordance, accuracy, sensitivity, stability, and species specificity.
Main Results:
- Complete genotypes were obtained from ≥100 pg of input DNA.
- High accuracy and specificity were achieved within 90 minutes.
- The method demonstrated reliability with challenging samples.
Conclusions:
- The developed method offers a rapid and cost-effective solution for InDel genotyping.
- This portable approach is suitable for personal identification and paternity testing.
- The technology addresses the limitations of current forensic DNA analysis techniques.
