Nguyen Quang, McKinney Jason, Johnson Donald J, Roberts Katherine A, Hardy Winters R
Los Angeles Police Department, Scientific Investigation Division, Los Angeles, CA 90032, USA.
J Forensic Sci. 2012 Jul;57(4):887-99. doi: 10.1111/j.1556-4029.2012.02106.x. Epub 2012 Apr 6.
In this proof-of-concept study, high-resolution melt curve (HRMC) analysis was investigated as a postquantification screening tool to discriminate human CSF1PO and THO1 genotypes amplified with mini-STR primers in the presence of SYBR Green or LCGreen Plus dyes. A total of 12 CSF1PO and 11 HUMTHO1 genotypes were analyzed on the LightScanner HR96 and LS-32 systems and were correctly differentiated based upon their respective melt profiles. Short STR amplicon melt curves were affected by repeat number, and single-source and mixed DNA samples were additionally differentiated by the formation of heteroduplexes. Melting curves were shown to be unique and reproducible from DNA quantities ranging from 20 to 0.4 ng and distinguished identical from nonidentical genotypes from DNA derived from different biological fluids and compromised samples. Thus, a method is described which can assess both the quantity and the possible probative value of samples without full genotyping.
在这项概念验证研究中，对高分辨率熔解曲线（HRMC）分析进行了研究，将其作为一种定量后筛选工具，用于在存在SYBR Green或LCGreen Plus染料的情况下，区分用微型短串联重复序列（mini-STR）引物扩增的人CSF1PO和THO1基因型。在LightScanner HR96和LS-32系统上对总共12种CSF1PO和11种HUMTHO1基因型进行了分析，并根据它们各自的熔解曲线进行了正确区分。短串联重复序列（STR）扩增子的熔解曲线受重复次数影响，单源和混合DNA样本还可通过异源双链体的形成进行区分。熔解曲线显示出独特性且在20至0.4 ng的DNA量范围内具有可重复性，并且能够区分来自不同生物体液和受损样本的DNA中相同与不同的基因型。因此，本文描述了一种方法，该方法无需进行全基因分型即可评估样本的数量和可能的证据价值。
