Evaluating the Efficiency of the Use of the Qiagen® QIAsymphony® with High Throughput Y-screening as an Alternative to Conventional Serology

As submitted by the proposer: Sexual Assault Kit backlogs have been a common topic in news stories in recent years throughout the United States. According to the Rape, Abuse and Incest National Network (RAINN), there are 237,868 victims of rape/sexual assault per year. The goal of this project is to develop a high throughput screening method for the presence of male DNA. The aim of this project is to increase laboratory efficiency by performing a Y-screen utilizing the Qiagen® QIAsymphony® on a phosphate buffered saline (PBS) extract instead of completing conventional serology. Historically, serology screening has been the method for processing sexual assault kits. Drawbacks of this method include: false positives and negatives, difficulty in automation, subjective interpretation, and lengthy time commitments to a microscope. Screening for male DNA (Y-screen) enables the process to be automated, using a 96 well format allowing for the screening of up to 88 samples simultaneously on a PCR-based method. This method brings objectivity to the screening process and provides a downstream correlation similar to the current DNA analysis techniques. A process unique to this approach is taking a larger cutting from the evidence; cells will be extracted from the sample with PBS buffer. One-tenth of this will be extracted utilizing DTT based extraction buffer on the Qiagen® QIAsymphony® that will lyse all cells. Quantitative PCR with simultaneous male and human DNA quantitation setup will then occur on the Qiagen® QIAgility® robotic liquid handler. Then the screening of samples based on the presence or absence of male DNA will occur. For positive samples, a differential separation and extraction will then be performed on the remaining original PBS sample extract. The cutting size and PBS soak allow for a homogenous sampling creating an accurate representation of the DNA on the evidence. This project will compare and evaluate laboratory efficiency of the Y-screen model versus conventional serology. All samples will be screened with both procedures. Analyst time and reagent use will be logged for the cost analysis. This project contains a research and/or development component, as defined in applicable law. ca/ncf