칸디다혈증 환자에서 분리된 Candida albicans 균주의 중합효소연쇄반응 및 Southern 교잡법을이용한 형별 분석
DNA Fingerprinting of Candida albicans Strains Isolated from Candidemic Patients by Polymerase Chain Reaction and Southern Hybridization Methods
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목 적 : 최근 Candida albicans에 의한 감염의 분자역학을 이해하기 위해 다양한 균 형별 검사가 시도되고 있으나, 가장 적합한 방법은 아직 정립되어있지 않다. 본 연구는 칸디다혈증 환자의 임상검체에서 분리된 C. albicans 균주들을 대상으로 두 가지 DNA fingerprinting법을 이용하여 균 분류를 시도하고 그 성적을 비교하여 보았다. 재료 및 방법 : C. albicans 칸디다혈증으로 진단된 환자 14명의 각종 임상검체(혈액 14주, 카테터 7주, 요 4주 및 호흡기 4주)에서 분리된 C. albicans 29주를 대상으로 하였다. Interrepeat (IR) PCR을 이용한 분석에는 primer 1245 및 1246의 두 가지를 이용하였고, C1 fingerprinting법은 염색체 DNA를 EcoRI으로 분해한 후 Ca3의 C1 단편을 probe로 이용하여 Southern 교잡법으로 실시하였다. 결 과 : 14명에서 분리된 총 29주는 IR PCR를 이용한 분석 결과, primer 1245에 의해 9가지 형, primer 1246에 의해서는 7가지 형, 두 primer를 조합할 경우 13가지 형으로 구분된 반면, C1 fingerprinting법에 의해 16가지 형으로 구분되었다. 두 가지 방법의 결과를 조합할 때 29주는 총 16가지 형으로 구분되어 C1 fingerprinting법을 단독으로 시행한 성적과 일치하였다. 두 가지 방법 모두에 의해 10명의 혈액과 기타 검체에서 분리된 균주는 환자별로 동일한 유형으로 구분된 반면, 1명에서 분리된 균주는 서로 다른 유형이었다. 반면, 3명에서 분리된 균주는 환자별로 모두 동일한 IR PCR 유형을 보였으나, C1 fingerprinting법에 의해서는 다르거나(1명), 유사한(2명) 유형을 보였다. 두 가지 방법을 조합하여 판정할 때 각 환자의 혈액에서 분리된 균주는 대부분 카테터(7명중 6명), 뇨(4명 중 4명) 혹은 호흡기(4명 중 3명)에서 분리된 균주와 유사하거나 동일한 유형이었다. 결 론:C. albicans 균주의 형별 분석에는 IR PCR법에 비해 C1 fingerprinting법이 분별력이 더 우수한 방법으로 생각되었으며, 칸디다혈증 환자의 카테터, 요 혹은 호흡기 검체에서 분리되는 균주는 대부분 혈액에서 분리된 균주와 동일하거나 유사한 유형을 보임을 알 수 있었다.
Abstract
Background:Although several molecular typing methods have been used to investigate C. albicans infections, there remains no "gold standard" method by which relatedness of C. albicans strains is determined. In this study, two DNA fingerprinting methods were compared for genotyping of clinical strains of C. albicans isolated from candidemic patients. Materials and Methods:Twenty-nine strains of C. albicans isolated from various clinical specimens (14 from blood, 7 from catheter, 4 from respiratory tract secretion, and 4 from urine) of 14 candidemic patients were analyzed. Primer 1245 and 1246 were employed for IR PCR and Southern blot hybridization method was used for C2 fingerprinting, with Ca3 and C1 as primers, after the fragmentation of DNA with EcoR1 Results:IR PCR method separated 29 isolates into 9 (1245 primer), 7 (1246 primer) and 14 (combination of two primers) types, whereas C1 fingerprinting identified 16 different types. By combining the IR PCR and C1 fingerprinting methods, total of 16 different genotypes were identified among 29 isolates from 14 patients, which is the same result obtained by the C1 fingerprinting only. Using both methods, blood and non-blood isolates from each patient produced identical genotypes for 10 patients and different genotypes for 1 patient. In three patients, isolates from blood and other site of each patient showed identical patterns by IR PCR fingerprinting, but appeared different (n=1) or similar (n=2) by C1 fingerprinting. Overall, for 87% (13/15) of patients, isolates collected from catheter (6 of 7 patients), urine (4 of 4 patients), or respiratory (3 of 4 patients) were identical or similar to the corresponding blood isolates. Conclusion:Our study shows that C1 fingerprinting method is more discriminatory than IR PCR for the molecular typing of C. albicans isolates. For the majority of patients, blood and other site isolates had identical or similar genotypes.
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