다중 중합효소연쇄반응을 이용한 Candida 균종동정
Identification of Candida Species by Multiplex Polymerase Chain Reaction
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배경：신속한 Candida 균종동정을 위하여 중합효소연쇄반응(polymerase chain reacation, PCR)에 기초한 방법들이 시도되고 있다. 본 연구에서는 Candida의 internal transcribed spacers와 topoisomerase II 유전자를 사용하여 Candida 균종동정을 위한 다중 PCR을 개발하고자 하였다. 방법：다중 PCR을 위하여 Dual Specificity Oligo (DSO) 시발체를 제작하였고, 8종류의 표준균주(C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae, C. dubliniensis)와 96균주의 임상분리 Candida spp. (C. albicans 51균주, C. parapsilosis 10균주, C. glabrata 10균주, C. tropicalis 9균주, C. krusei 6균주, C. guilliermondii 5균주, C. lusitaniae 5균주)를 대상으로 다중 PCR 후의 아가로오스 겔 전기영동으로 균종을 동정하였다. 결과：DSO 시발체를 사용한 다중 PCR로 8종류의 표준균주의 감별이 용이하였다. 임상분리 96균주 모두 전통적인 방법으로 동정된 균종과 동일한 균종으로 확인되었다. 결론：Candida 균종동정 시 DSO 시발체를 사용한 다중 PCR은 일반 검사실에서 간편하고 신속하게 시행하기에 유용한 방법으로 사료되었다.
Abstract
Background: Polymerase chain reacation (PCR)-based methods have been described for rapid detection and identification of Candida spp. Multiplex PCR assay was developed using internal transcribed spacers and topoisomerase II gene for the accurate identification of Candida species. Methods: We designed Dual Specificity Oligo (DSO) primers for multiplex PCR. Multiplex PCR was followed by agarose gel electrophoresis to test 8 type strains (C. albicans, C. arapsilosis, C. glabrata, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae, C. dubliniensis) and 96 clinical isolates (C. albicans 51 isolates, C. parapsilosis 10 isolates, C. glabrata 10 isolates, C. tropicalis 9 isolates, C. krusei 6 isolates, C. guilliermondii 5 isolates, C. lusitaniae 5 isolates) of Candida spp. Results: With multiplex PCR using DSO primers, the eight Candida type strains each could be easily differentiated and all 96 clinical isolates were identified as the same species as were identified by the conventional method. Conclusion: Multiplex PCR followed by electrophoresis can be useful for the simple and rapid identification of Candida species in routine laboratories.
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