I have previously asked a more general question about how PCR sequencing reaches all possible DNA targets here. I want to ask more specifically now: what evidence is there that common PCR DNA purification kits can isolate DNA from pathogens engulfed in macrophages?

Specifically what such a KIT would have to achieve is to destroy the double lipid cell membrane of a macrophage and destroy the cell walls of engulfed bacteria or fungi. (The latter of which even contain chitin)

I am looking for as specific evidence as possible proving that if one takes a whole blood sample and employs a purification KIT, any microorganism's DNA engulfed in a macrophage should be isolated. Please allow me to say that I am looking for evidence beyond just the manufacturers specifications. I would like to find publications, book chapters or other written statements that specifically show and discuss how isolation techniques work on the barriers I mentioned or even better how they work on intra-cellular micro organisms. Thank you!

  • $\begingroup$ What do you mean by a "PCR DNA purification kit" ? Can you link to such a kit? $\endgroup$ – March Ho Nov 3 '15 at 12:42
  • $\begingroup$ Also, colony PCRs routinely work on either colonies of bacteria or isolated mammalian cell culture, so there is no reason bacteria inside mammalian cells won't work. I can't say the same for fungi, since I have not worked with yeast. $\endgroup$ – March Ho Nov 3 '15 at 12:45
  • $\begingroup$ @March Ho: the link you asked for, qiagen.com/de/shop/sample-technologies/dna-sample-technologies/… $\endgroup$ – B M Nov 3 '15 at 19:51
  • $\begingroup$ @March Ho: so your reasoning ist that since isolation techniques work on mammalian cells and on bacteria, they should also work on one inside the other? That might be the case but does not follow by pure logic! The kit you use on one (bacteria/mammalian cells) might not work on the other and even if you use the same kit for both it might not isolate the dna of bacteria inside mammalian cells for reasons of: too short exposure time or not enough reactants or competitive binding or unforseen reactions or altered cell wall of intracellular bacteria or... $\endgroup$ – B M Nov 3 '15 at 20:02
  • $\begingroup$ I didn't use a kit, but merely ran a PCR using the cells instead of purified DNA as the template. $\endgroup$ – March Ho Nov 3 '15 at 21:16

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