Use of macroporous polypropylene filter to allow identification of bacteria by PCR in human fecal samples.
The detection of pathogenic bacteria directly in human fecal specimens by PCR, requires removal of PCR-inhibitory substances. To investigate whether five different macroporous filters (polypropylene, nylon, polyester, polyethylene, fluorocarbon) could retain polysaccharides, major PCR inhibitors, an in vitro model and human fecal samples were used. The in vitro model consisted of Xanthum gum solutions (3 mg/ml PBS), a bacterial polysaccharide, to which Helicobacter pylori cells were added. Fecal samples from healthy volunteers were spiked with H. pylori and Mycobacterium paratuberculosis cells. Polysaccharide concentrations were significantly reduced only by the polypropylene but not by the other filters. Accordingly, both Xanthum gum solutions and spiked fecal specimens became PCR positive only after filtration with the polypropylene filter. We conclude that this filter can be used to prepare a bacterial DNA template suitable for PCR analysis from human feces.
Cavallini
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Laboratory of Biochemistry, I.R.C.C.S. S. de Bellis, Scientific Institute for Digestive Diseases, Castellana Grotte (BA), Italy.
Notarnicola
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M
Berloco
P
P
Lippolis
A
A
De Leo
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D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D005243:Feces / Q000737:chemistry / Q000382:microbiology*; D005374:Filtration / Q000295:instrumentation; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D011126:Polypropylenes; D011135:Polysaccharides, Bacterial
D004269:DNA, Bacterial; D011126:Polypropylenes; D011135:Polysaccharides, Bacterial; C002563:xanthan gum