PATHOGEN DETECTION AND QUANTIFICATION CORE

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The pathogen detection and quantification core (PDQC) was established in late 2003 following the approval of the program during the base grant review process. This core is designed to supply testing for the specific pathogen free colonies. The core is currently functional. Diagnostic tests for SIV, STLV and Herpes B infections are currently provided. For SIV and STLV we are also providing confirmation assays. The algorithm for herpes B involves screening by the PDQC followed by confirmation in the National Reference laboratory. New diagnostic tests are being developed and standardized for SRV. Since testing different peptide formats did not offer a good sensitivity, we are also optimizing PCR screening for SRV. The viral diagnostic core processed and tested more than 5,000 samples in 2005 and over 6,000 in 2006 and 2007. A new strategy for economical sample processing is in progress. In 2008 over 10,000 clinical tests were provided for the animals in the TNPRC colony We are currently exploring new strategies for B virus and SRV diagnostic. It is expected that by the end of 2009 routine testing will be performed through the Luminex multiplex bead technology. Real-time PCR is offered as a part of capabilities of the PDQC. The major objectives are: 1. to assist PIs using the facility. In this respect, all of the investigators previously using the rt-PCR core received the needed support. New investigators were helped to implement their procedures. 2. To provide service for viral quantification. We optimized a rt-PCR assay for the SIVmac/SHIV quantification. This new assay is currently under optimization. We provide quantification assays for SIVagm and SIVrcm. We also provide genequantification (CCR5, TGF-beta, GATA-3, IL6, IFN-gamma). A new 7900 Squence amplifier was purchased in 2008. Real-time PCR quantification was provided in 2008 for 8 projects. Over 1000 assays have been carried out on both plasma as well as tissues in 2008.