Validation of Multiplex Polymorphic STR Amplification Sets Developed for Personal Identification Applications

    Volume 41, Issue 4 (July 1996)

    ISSN: 0022-1198

    CODEN: JFSOAD

    Page Count: 9


    Lins, AM
    R & D Scientists, Director, Promega Corporation, Madison, WI

    Koons, BW
    Research Chemist, Chemist, Forensic Science Research and Training Center, FBI Academy, Quantico, VA

    Sprecher, CJ
    R & D Scientists, Director, Promega Corporation, Madison, WI

    Comey, CT
    Research Chemist, Chemist, Forensic Science Research and Training Center, FBI Academy, Quantico, VA

    Crouse, C
    Serology/DNA Supervisor, Palm Beach Sheriff's Crime Lab, West Palm Beach, FL

    Endean, D
    V. P. and Product Development Technologist, Blood Center of Southeastern Wisconsin, Inc., Milwaukee, WI

    Schumm, JW
    R & D Scientists, Director, Promega Corporation, Madison, WI

    Micka, KA
    R & D Scientists, Director, Promega Corporation, Madison, WI

    Pirelli, K
    V. P. and Product Development Technologist, Blood Center of Southeastern Wisconsin, Inc., Milwaukee, WI

    Ma, M
    Research supervisor and Criminalist, California Department of Justice, DNA Laboratory, Berkeley, CA

    Duda, N
    Research supervisor and Criminalist, California Department of Justice, DNA Laboratory, Berkeley, CA

    Lee, SB
    Research supervisor and Criminalist, California Department of Justice, DNA Laboratory, Berkeley, CA

    Abstract

    Polymorphic short tandem repeat (STR) loci, which typically consist of variations in the number of 3–7 base pair repeats present at a site, provide an effective means of personal identification. Typing can be accomplished by amplification of genomic DNA using the polymerase chain reaction (PCR) and locus-specific primers, separation of amplified alleles using gel electrophoresis and their display using silver staining or fluorescent detection. Primers for several STR loci can be combined in a single multiplex reaction so typing of multiple loci can be accomplished rapidly and with less DNA than required if each locus were analyzed separately. Before such multiplex systems are used in forensic or paternity applications, it is desirable that they undergo testing for their reliability.

    This study evaluates the performance of two STR triplex systems, one containing the loci HUMCSF1PO, HUMTPOX, and HUMTH01, and the other containing HUMHPRTB, HUMFESFPS, and HUMVWFA31. Protocols for amplification of these two triplexes, and their corresponding monoplexes, were evaluated for sensitivity of detection, resistance to changes in the annealing temperature of the amplification protocol, and the ability to identify the minority contributor in amplification of mixed samples. In addition, five laboratories determined the alleles of twenty DNA samples, each extracted by one of four different extraction methods. The results illustrate that the two STR triplex systems and the monoplex systems contained within them can be used with as little as 0.25 ng of DNA template. Both triplexes amplified with 100% success using the Perkin Elmer Model 480 thermal cycler. With the GeneAmp 9600 System, the CTT triplex amplified with 100% success and the HFv triplex in 95.6% of attempts. These experiments meet many requirements for use in validation of DNA typing systems for forensic cases and paternity identification.


    Paper ID: JFS13959J

    DOI: 10.1520/JFS13959J

    ASTM International
    is a member of CrossRef.

    Author
    Title Validation of Multiplex Polymorphic STR Amplification Sets Developed for Personal Identification Applications
    Symposium , 0000-00-00
    Committee E30