Development and validation of the 6-dye lyophilized fluorescent multiplex PCR system

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Authors

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Publication Details

Journal
PLoS ONE
Published
2026-08-27
DOI
https://doi.org/10.1371/journal.pone.0356258
Primary Topic
Forensic and Genetic Research
Type
article
Field-Weighted Citation Impact
0.00

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article

Development and validation of the 6-dye lyophilized fluorescent multiplex PCR system

Lina Jia, Cui Yuling, Ming Ji, Jing Wang et al.
PLoS ONE
Forensic and Genetic Research
article

Development and validation of the 6-dye lyophilized fluorescent multiplex PCR system

Lina Jia, Cui Yuling, Ming Ji, Jing Wang, Zhe Zhang, Wen Song, Hao Yu, Lei Shang, Xue Bai
article en

Abstract

Aiming to address the issues associated with traditional liquid STR reagents-such as dependence on cold-chain storage, cumbersome operation, and unsuitability for on-site rapid detection-this study has successfully developed a six-color fluorescent lyophilized multiplex amplification reagent. The reagent includes 29 autosomal STR loci and 1 gender identification locus. By screening high-tolerance Taq polymerase, optimizing lyoprotectant formulations, and developing a lyophilization process based on thermodynamic analysis, stable ambient-temperature storage of key reagent components has been achieved. Performance validation demonstrates that the lyophilized reagent exhibits high sensitivity (100% detection at 0.125 ng DNA), remains stable for 60 days at room temperature, rehydrates rapidly (<1 second), and shows some tolerance to common inhibitors. Benefiting from the "whole-volume loading" advantage, its detection capability for trace samples surpasses that of traditional liquid reagents. In parallel comparison with similar liquid reagents, it achieves completely consistent locus detection rates (100%), fully meeting the requirements for individual identification. With simple operation and a ready-to-use, disposable design, this reagent effectively overcomes the reliance on cold-chain logistics. It provides robust technical and reagent support for rapid on-site forensic testing, emergency response, grassroots applications, and DNA database construction, demonstrating significant application prospects.

PLoS ONEVol. 21(8)
Hebei Medical University (CN), Ministry of Public Security of the People's Republic of China (CN), Institute of Forensic Science (CN)
Special Funds for the Basic Research and Development Program in the Central Non-profit Research Institutesof China, Institute of Forensic Science, Ministry of Public Security
Gender equality
Openalex Percentile: Top 11%
Forensic and Genetic Research
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