NOTE: urine sample contains human epithelial cells

​Pre-incubation of lysates containing human urine with 100 cfu / sample of E. coli with PCRopsis Cell for just 10 minutes allows for bacterial detection through direct PCR and amplification of a bacterial 16S rRNA fragment to a level that is comparable to Qiagen's QIAamp® DNA Kit. No amplification was observed when the lysate was directly placed into the PCR mixture without pre-incubation with PCRopsis Cell or DNA isolated using QIAamp® DNA Kit. 
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Note: PCR Study used PCRopsis 5x PCR Master Mix (cat. no. PMM001)

​Pre-incubation of lysates containing human urine with 100 or 1000 cfu / sample of E. coli with the Cell PCRopsis tubes for just 10 minutes allows for bacterial detection through direct real-time PCR and amplification of a bacterial 16S rRNA fragment to a level that is comparable to Qiagen's QIAamp® DNA Kit. No amplification was observed when the lysate was directly placed into the PCR mixture without pre-incubation with Cell PCRopsis tubes or DNA isolated using QIAamp® DNA Kit. Real-time PCR utilized 1 uL of lysate or isolated DNA, and SYBR green for detecting the amplified bacterial 16S rRNA fragment.

Statistical Analysis (qPCR results: Cell PCRopsis vs QIAamp® DNA Kit):

1. Kruskal-Wallis 1-way ANOVA (P<.05):  no significant difference
2. Dunn's Multiple Comparison Test (all pairwise combinations, P<0.05):  no significant difference

These results demonstrate that thermal lysis of E. coli followed by processing with Cell PCRopsis results in statistically comparable amplification of a bacterial 16S rRNA DNA fragment when compared to Qiagen's QIAamp® DNA Kit. It's possible that other microorganisms may require pre-incubation with a detergent and / or treated with proteinase K to achieve comparable cell lysis and DNA amplification.
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Note: qPCR Study used PCRopsis 5x PCR Master Mix (cat. no. PMM001)