The objective of our NIAID Biodefense Phase I SBIR program was to develop, and commercialize field deployable nucleic acid analysis devices for use in biodefense applications. We have begun to sell IsoAmp II kits that will allow our clients to develop home brew assays (we list customers in the commercialization plan). For example, the LANL has obtained NIH funding (R01) to develop an assay system for detecting influenza virus using our IsoAmp II assay formulation (www.lanl.gov/science/1663/flu.php). These kits contain an enhancer protein, discovered in our Phase I, that accelerates helicase dependent amplification (HDA);i.e., a thermostable single strand DNA binding protein. HDA chemistry now allows for the specific amplification of both ribonucleic acids (RNA), and deoxyribonucleic acids (DNA). Moreover, we have optimized our primer design process to facilitate assay development. We also collaborated with the Keck Graduate Institute (KGI) to develop a small, portable nucleic acid analysis device (a.k.a. Rabbit). Finally, we have developed a number of DNA probe based assays for biodefense targets;e.g. Bacillus anthracis and Ebola virus. In this Phase II, we propose to put into place a functioning QMS at BioHelix, manufacture cGMP reagents and devices that perform the our assays, and to validate our assay system for typing HSV-1 &HSV-2 with a large number (~1,000) clinical samples. The data generated from this clinical study will be used to apply for regulatory clearance from FDA for the sale of this diagnostic system. We have selected HSV as a target for this first clinical assay because this virus is the lead cause of sexually transmitted disease (STD) in the United States. In addition, molecular testing for HSV DNA has greater sensitivity than the traditional viral culture (sensitivity of more than 95 percent, compared with 75 percent for culture). Despite this fact, molecular tests are not as widely used as culture for the diagnosis of genital herpes because of the higher cost of molecular tests. Moreover, molecular testing is viewed as a complex process requiring the isolation of DNA from clinical samples. Finally, as there are no FDA cleared commercial molecular assays for HSV, implementing such tests in clinical laboratories is burdensome. We believe we can offer a solution to this problem. Genital and oral swab specimens collected in Cellmatics Transport Kit buffer (Becton Dickinson, Sparks, MD) can be diluted 1/2000 for direct analysis by the IsoAmp HSV assay. This dilution based sample processing will greatly reduce the degree of complexity of our assay system. In addition, we have begun to collaborate with GE Healthcare to develop a Ready-To-Go(tm) IsoAmp formulation for our assays that will eliminate the need for multiple, precise volumetric measurements. BioHelix and GE Healthcare plan to develop a formulation under design control in order to manufacture it under cGMP. The instrument developed by KGI will also be transferred to a cGMP manufacturer (Source Scientific), such that by mid-2009 we expect to have a cGMP assay system ready for clinical validation studies.
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