ứng dụng kỹ thuật RFLP-PCR phát hiện nấm Cryptococcus neoformans gây bệnh ở người

Cryotococcus neoformans sample was collected from cerebrospinal fruild to extract DNA and perform PCR in the same condition. PCR products was subjected to separate restriction digestions with MspI enzyme for identification. PCR and restrictive products was compered by 100bp marker. The results showed that RFLP-PCR method with ITS1, ITS4 primers and MspI enzyme allows simple, rapid and reliable to identify C. neoformans with 1.5mM Mg2+, 150uM of each deoxyribonucleotide, 0.2-0.3pmol/ul of each primer and annealing temperature 55°C. The limited concentration of the detection of C. neoformans was less than 100 yeast cells per ml.