Detection of Cryptococcus neoformans Gene Using PCR

Abstract

We studied detection of the Cryptococcus neoformans gene using PCR method. The primer was designed from 20 mer of Cryptococcus neoformans URA 5 gene. Conditions of the PCR method were denaturing at 94°C for 60sec, annealing at 63°C for 90sec, extension 72°C for 60sec, and this was repeated for 36 cycles. DNA extraction was conducted by the glass bead method, the enzyme method or a combination of the two and the glass bead method showed the highest extraction rate in experiments. The series of experiments revealed the high specificity of the adapted PCR method. Sensitivity in nested PCR was 100pg in quantity of DNA and 1×10³ in diluted solution.