Restriction Fragment Length Polymorphism and Polymerase Chain Reaction-HLA DQa Analysis of Casework Urine Specimens
Medintz, I
Research Associate, Research Associate, and Research Assistant, respectively, John Jay College of Criminal Justice, City University of New York, Department of Forensic Sciences, New York, NY.

Chiriboga, L
Research Associate, Research Associate, and Research Assistant, respectively, John Jay College of Criminal Justice, City University of New York, Department of Forensic Sciences, New York, NY.

McCurdy, L
Research Associate, Research Associate, and Research Assistant, respectively, John Jay College of Criminal Justice, City University of New York, Department of Forensic Sciences, New York, NY.

Kobilinsky, L
2Prof. Biology and Immunology, John Jay College of Criminal Justice and Member, Doctoral Faculty, Ph.D. Program in Biochemistry, Graduate Center, City University of New York, New York, NY.


Abstract
DNA was isolated from casework urine samples previously submitted for toxicological analysis. The quality and quantity of DNA isolated was determined by spectrofluorometry and agarose yield gel electrophoresis. Hae III restricted samples were then resolved by analytical agarose gel electrophoresis, transferred to a membrane by Southern blotting and hybridized with a chemiluminescently-labelled (D2S44) probe. The DNA fragment banding patterns were indistinguishable from the DNA banding patterns of blood specimens collected from the same donor. Only 5 of 20 samples yielded banding patterns and the banding intensity relative to background was low. Genomic DNA was also obtained from casework samples by Chelex extraction, amplified by polymerase chain reaction (PCR) and then genotyped for human leucocyte antigen (HLA) DQa. Of 20 specimens, 13 (65%) were typed correctly producing identical results for urine and blood specimens obtained from the same donor. Aging studies of casework samples and normal samples (from a non-drug using population) were also conducted with PCR-HLA DQa analysis. Results of these studies indicate that amplification by PCR was more likely to produce positive results. Based on these findings, we conclude that PCR-initiated analysis is more suitable than RFLP analysis for individualization of urine samples.

Keywords:
DNA, forensic science, HLA DQa, polymerase chain reaction, polymerase chain reaction (PCR), restriction fragment length polymorphism, RFLP, toxicology, urine

Paper ID: JFS396941372

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Author Medintz I, Chiriboga L, McCurdy L, Kobilinsky L Title Restriction Fragment Length Polymorphism and Polymerase Chain Reaction-HLA DQa Analysis of Casework Urine Specimens Symposium , Committee on