AWWA WQTC69415

AWWA WQTC69415 Development of "Bioball" as Internal Control for Detection of Helicobacter pylori by Quantitative PCR
Conference Proceeding by American Water Works Association, 11/01/2008

Sen, Keya

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This powerpoint presentation begins by providing a brief overview of the process of detection for bacteria(sample collection to final detection). Development of qPCR for H. pylori is presented, along with controls needed, and possible molecules for positive control (IC). Chosen strategy included:modified DNA fragment with change in the probe region- used in a duplex reaction; and, PCR mutagenesis used to modify target ureAfragment by four bases. Result 1:if low numbers of H. pylori to be detected then nomore than 10-50 molecules IC can be added in a Duplex PCR Assay. Result 2:E.coli pKS10 with the modified fragment isrecovered as well as H.pylori from distilled water;therefore, E. coli pKS 10 can be used as matrixspike in a field sample; detected in a duplex qPCR assay with modified fragment being detected by probe 2 (VIC); and, target H. pylori probe 1 (Fam).Result 3: recovery of Bioball comparable to fresh cells of E. coli; can be used to spike water in place of fresh E. coli; and, use a Bioball containing 30 cells. Results 4: No H. pylori detected from the water utilities; turn around time 6-8 hrs.; one sample was inhibited slightly, as indicated from the recovery of Bioballs spiked H. pylori recovered 99% efficiency from these samples, except for the one inhibitory; four of the source waters were strongly inhibitory; and, in addition, tested two wastewater samples. Includes tables, figures.