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METHODS FOR MEASURING POLYMERASE ACTIVITY USEFUL FOR SENSITIVE, QUANTITATIVE MEASUREMENTS OF ANY POLYMERASE EXTENSION ACTIVITY AND FOR DETERMINING THE PRESENCE OF VIABLE CELLS
METHODS FOR MEASURING POLYMERASE ACTIVITY USEFUL FOR SENSITIVE, QUANTITATIVE MEASUREMENTS OF ANY POLYMERASE EXTENSION ACTIVITY AND FOR DETERMINING THE PRESENCE OF VIABLE CELLS
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机译:测量任何聚合酶延伸活性的敏感,定量的测量和确定活细胞存在的聚合酶活性的方法
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摘要
A novel, highly sensitive, quantitative and rapid DPE-PCR assay is disclosed that can be used to enumerate prokaryotic cells when presenting a purified or selected cell type and that has the capability to reproducibly measure DNA polymerase extension activity from less than 10 cfu of bacteria via coupling to bead lysis. Also disclosed is the potential for the DPE-PCR assay of the invention to universally detect microbes by testing a panel of microorganisms comprised of gram-negative bacteria, gram-positive bacteria and Candida species. Furthermore, it is disclosed that the DPE-PCR assay of the invention can be used to assess bacterial cell viability, provided via the reproducibly strong correlation between DNA polymerase extension activity and proliferation as indicated by the presence of cfu. It is believed that the disclosed assay of the invention can be a useful quantitative tool for a wide range of testing applications within pharmaceutical, environmental, food and clinical settings.
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