The point mutation of cytochrome P-450 lanosterol 14-? demethylase ERG11 gene in Fluconazole-resistant Candida albicans / 中华传染病杂志

ABSTRACT

Objective To investigate the point mutation of the open reading frame of cytochrome P-450 lanosterol 14-? demethylase gene ERG11. Methods In order to identify such alterations, the DNA was extracted by enzyme lysis methods and the PCR was performed. The PCR products were purified and cloned into PBS vector, then transformed into DH5? and sequenced. Results Twenty nine point mutations were identified in this 15 resistant isolates, most of which were different from susceptible strains. The mutations included 12 missense substitutions: F72L, D81G, D116E, K128T, Y132H, E266D, D294G, S361P, M374V, P386L, H400R, and Q474K, of which 6 had not been described previously (D294G, S361P, M374V, P386L, H400R, and Q474K). The other mutations included a frameshift, and 17 silent mutations. Conclusions The point mutation of resistant isolate is different from that of the susceptible isolate. It is suggested that the drug resistance is related with by the newly found alterations, including D294G, S361P, M374V, P386L, H400R, Q474K, and a frameshift.