Abstract
In this paper, we demonstrate a rapid analysis procedure for use with a small set of rapidly mutating Y chromosomal short tandem repeat (Y-STR) loci that combines both rapid polymerase chain reaction (PCR) and microfluidic separation elements. The procedure involves a high-speed polymerase and a rapid cycling protocol to permit PCR amplification in 16 min. The resultant amplified sample is next analysed using a short 1.8-cm microfluidic electrophoresis system that permits a four-locus Y-STR genotype to be produced in 80 s. The entire procedure takes less than 25 min from sample collection to result. This paper describes the rapid amplification protocol as well as studies of the reproducibility and sensitivity of the procedure and its optimisation. The amplification process utilises a small high-speed thermocycler, microfluidic device and compact laptop, making it portable and potentially useful for rapid, inexpensive on-site genotyping. The four loci used for the multiplex were selected due to their rapid mutation rates and should proved useful in preliminary screening of samples and suspects. Overall, this technique provides a method for rapid sample screening of suspect and crime scene samples in forensic casework.
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Abbreviations
- FTA® :
-
Fast technology for analysis of nucleic acids
- RFU:
-
Relative fluorescence units
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Acknowledgments
The authors would like to thank the Streck and Agilent Technologies for their technical assistance and support. We also thank Fabiana Taglia for technical assistance. The authors gratefully acknowledge the donation of equipment from the Agilent Technologies and Streck. This project was partially supported by Award No. 2015-IJ-CX-K038 of the National Institute of Justice, Office of Justice Programs, U.S. Department of Justice. Points of view in the document are those of the authors and do not necessarily represent the official view of the U.S. Department of Justice.
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Sampling of individuals was done with their informed consent, via signed consent forms, and in accordance with the guidelines and regulations set in place by the FIU IRB board and approved under the reference number 101831.
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Gibson-Daw, G., Albani, P., Gassmann, M. et al. Rapid microfluidic analysis of a Y-STR multiplex for screening of forensic samples. Anal Bioanal Chem 409, 939–947 (2017). https://doi.org/10.1007/s00216-016-9950-9
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DOI: https://doi.org/10.1007/s00216-016-9950-9