Rapid PCR based detection methods for the fungal species from mycotic keratitis patients. (Fungal identification upto the species level or at least genetic level)
Sampling methods from the patients for the PCRR and the quantity of sample required and all.
What are the PCR primers used for the detection of common fungal species isolated from mycotic keratitis (Aspergillus, Fusarium, UIH, UID, Curvularia, Bipolaris, Exerohilum, etc)
Thank you for your effort on my query
Dear colleague,
This paper described rapid identification of a dematiaceous fungus causing corneal infection by PCR. DNA was extracted directly from the infected tissue: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC165366/
Those authors used universal primers for the fungal ITS region. This locus is the oficial barcode for fungi and provides enough information for species or species-complex identitification in most cases.
Success!
Hugo
Dear Mr. Hassan
detection by PCR and by ITS2 and 5.8S Ribosomal DNA Typing
please refere to the link
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC88253/
Dear colleague
I agree with two others colleagues. ITS primers are good enough for species or species complex identification (after sequencing your PCR products). My experience is on plant pathogens (and we used ITS1 and ITS4) but I think it doesn't change for yours, since we have also fungal plant pathogens as Fusarium, Aspergillus, Curvularia, Bipolaris...
Succes!
Beatrice
Dear Mr. Hassan !
You may use primer pair ITS1F and ITS4 or V9G and ITS4 which will cover entire ITS region (ITS1, 5.8S and ITS2), details you will find here in this article (https://goo.gl/cri7eb). These primers I used, were successful for environmental sample as well as culture mycelia.
For fungal identification I have used ITS primers. The best ones that I have utilized are the fungal specific primers NSI1F and NLB4R (Martin and Rygiewicz 2005).
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