With an history of Endophthalmitis, a patient’s Silicone oil (Retinal tamponade) was suspected for infectious condition and sent to laboratory. Gram’s smear shows occasional GPB. But PCR targeting 16s rRNA for Eubacteria genome is negative. Tried DNA spiking and found that no inhibitors present. What could be the reason??
What is GPB?(gram positive bacteria?)
Did you check quality and quantity of extracted DNA?
How did you confirm negative result for PCR product?
Did you try to culture?
Give more information please.
Regards.
I have no experience of this situation but if you thoroughly mix a sample of oil with water might the bacteria lyse and the dna would separate into the aqueous phase, A quick spin would separate the phases then pcr of the water phase ( nested pcr if necessary) seems a possible method
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