Demonstration of a Sample Preparation Method for Biological Detection Based on a Novel Membrane Fractionation Technology.

摘要

L WI funding has been obtained to gather proof-of-concept data for passing DNA from lysed bacteria through a membrane and for concentrating DNA in the concentrator cell. AlburtyLab has data showing high efficiency ( approx. 99%) for passing Bovine serum albumin through an appropriate membrane and data showing high efficiency (approx. 60%) for concentrating 0.03 liD microspheres with the system. Data showing similar efficiencies for DNA would significantly improve the quality of future SBIR submissions. AlburtyLab specifically requested this funding for the purpose of performing quantitative testing to determine the efficiency data for (1) passage of DNA through a cleanup InnovaPrep(copyright) membrane and (2) concentration of DNA in a final concentrating InnovaPrep membrane. AlburtyLab contracted with Midwest Research Institute (MRI), of Kansas City, Missouri, to prepare a stock of Bacillus globigii (Bg), American Type Culture Collection (ATCC) 9733, available from the Critical Reagents Program as ANG-BACI051, also known as Bacillus atrophaeus. MRI lysed the stock with a standard, commercially available bead beater (BioSpec Mini). The lysed stock was delivered to AlburtyLab for processing with the dual stage InnovaPrep system. All testing was performed by AlburtyLab personnel at the downtown engineering laboratory located in Drexel, Missouri. All resulting samples obtained after feed processing through the dual-stage InnovaPrep system were returned to MRI for analysis by quantitative PCR (qPCR). Detailed testing procedures are given in Section 2. Testing results are presented in Section 3. Section 4 contains discussion and conclusions. A total of 15 runs were conducted at two feed titer test levels. In the text, the individual concentrating cycles performed by the dual-stage InnovaPrep system are referred to as runs and the series of runs performed during a single feed titer level is referred to as a test. 1 LWI- 2008-.