One-Step Multiplex PCR for Simultaneous Detection and Identification of Eight Medically Important <i>Candida</i> Species
Fukatsu Akira, Osamu Tsuzukibashi, Hidenori Suzuk, Katsuhiro Asaka, Yoshinori Ono, Fuchigami Mana, Taira Kobayashi, Satoshi Uchibori, Yuji Takahashi, Komine Chiaki, Yoshimi Konishi, Yuki Ogura, Hiroko Omori, Masanobu Wakami, Hiroshi Murakami, Masahiko Fukumoto
Abstract
Recently, the incidence of Candida infections has substantially increased. Conventional identification methods for Candida species are technically difficult to conduct and cannot accurately distinguish each species. The purpose of the present study was to design primers to identify and detect simultaneously eight medically important Candida species using one-step multiplex PCR. PCR primers were designed based on partial sequences of intergenic spacer (IGS) and internal transcribed spacer (ITS) genes of eight medically important Candida species. These primers were able to distinguish each Candida species and did not display cross-reactivity with representative Candida species other than the eight Candida species. Moreover, our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and worked without requiring DNA extraction.