Keywords
DNA databank
PCR
dermatology
leprosy
How to Cite
Abstract
Skin biopsies from leprosy diagnostic services can be great sources of material for retrospective studies
concerning human and Mycobacterium leprae genetic. However, fixation and paraffin embedding procedures make difficult to obtain good quality DNA to PCR amplification. Thus, paraffin-embedded samples require special protocols to DNA extraction. The aim of this paper is to present an alternative method for DNA extraction based on the combination of heat and enzymatic digestion. The sections were heated at 120ºC at pH 9.0, submitted to enzymatic digestion with proteinase K and the DNA was extracted by using phenol:chlorophorm:isoamilic alcohol. PCR amplification for regions in TNF and LTA human genes was successful for 85.4 % of specimens. Regarding M. leprae DNA, we obtained amplification in 67.6%, 48.5%, 36.7% and 64.8% for the TA18, GTA9, TTC and RLEP markers, respectively. We conclude that this is
an inexpensive method which provided a satisfactory yield of a good quality DNA for PCR from paraffin- em-bedded skin biopsies.
References
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This work is licensed under a Creative Commons Attribution 4.0 International License.
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