Gibson-Daw Georgiana, Albani Patricia, Gassmann Marcus, McCord Bruce
Department of Chemistry and Biochemistry and International Forensic Research Institute, Florida International University, University Park, Miami, FL, 33199, USA.
School of Chemical Sciences, University of Auckland, 23 Symonds Street, Building 302, Auckland, 1010, New Zealand.
Anal Bioanal Chem. 2017 Feb;409(4):939-947. doi: 10.1007/s00216-016-9950-9. Epub 2016 Nov 30.
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. Graphical abstract ᅟ.
在本文中，我们展示了一种用于一小套快速突变的Y染色体短串联重复序列（Y-STR）基因座的快速分析程序，该程序结合了快速聚合酶链反应（PCR）和微流控分离元件。该程序涉及一种高速聚合酶和快速循环方案，以允许在16分钟内进行PCR扩增。接下来，使用一个1.8厘米的短微流控电泳系统对所得扩增样品进行分析，该系统可在80秒内产生四位点Y-STR基因型。从样品采集到得出结果，整个过程耗时不到25分钟。本文描述了快速扩增方案以及该程序的重现性、灵敏度及其优化研究。扩增过程使用小型高速热循环仪、微流控装置和紧凑型笔记本电脑，使其便于携带，并且可能有助于快速、廉价的现场基因分型。用于多重分析的四个基因座因其快速突变率而被选中，应证明对样品和嫌疑人的初步筛查有用。总体而言，这项技术为法医案件工作中嫌疑人及犯罪现场样品的快速样本筛查提供了一种方法。图形摘要ᅟ。
