Identification of Candida Species from Blood Cultures with Fluorescent In Situ Hybridization (FISH), Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Conventional Methods

Yıl 2010, Cilt: 2010 Sayı: 3, 183 - 191, 01.03.2010

Gülay Börekçi Gülden Ersöz Feza Otağ Hakan Özturhan Sebahat Şen Hilal Akalın Fatma Söylemez Yusuf Özkul Gürol Emekdaş

https://doi.org/10.5174/tutfd.2009.01700.1

Öz

Objectives: Rapid and accurate identification of Candida species from blood cultures is crucial to ensure effective antifungal therapy and to reduce the morbidity and mortality associated with bloodstream fungal infections. In this study, we aimed to identify Candida spp. from blood culture samples with fluorescent in situ hybridization (FISH), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and conventional methods. Materials and Methods: A total of 50 yeast positive samples out of 325 blood culture positive samples and 50 blood culture negative samples were examined by FISH, PCRRFLP and conventional methods to identify Candida spp. Results: All three methods generally were compatible for identification of single-species Candida spp. (p<0.001). But, FISH and PCR-RFLP for the identification of multi-species Candida spp. were found more compatible than conventional methods (p<0.001). Besides, FISH is cheaper and quicker than the other two methods in the identification of Candida spp. from blood culture positive samples. The rates of multi-species candidemia with FISH, PCR-RFLP and conventional methods were 20%, 6% and 4%, respectively. Conclusion: Both PCR-RFLP and FISH methods might be preferred for the rapid identification of Candida spp. from blood culture positive samples. However, FISH is a more suitable method for the detection of multi-species candidemia. Amaç: Kan kültürlerinden Candida türlerinin doğru ve hızlı identifikasyonu fungal kan dolaşımı infeksiyonlarla ilgili mortalite ve morbiditeyi azaltmak ve etkili antifungal tedavi sağlamak için önemlidir. Bu çalışmada, kan kültürü örneklerinden Candida türlerinin floresan in situ hibridizasyon (FISH), polimeraz zincir reaksiyonu-restriksiyon fragment uzunluk polimorfizmi (PCR-RFLP) ve konvansiyonel yöntemlerle identifikasyonu amaçlandı. Gereçler ve Yöntemler: Candida türlerininin identifikasyonu için 325 kan kültürü pozitif örnekten maya pozitif olan 50 örnek ile kan kültürü negatif 50 örnek FISH, PCR-RFLP ve konvansiyonel yöntemlerle incelendi Bulgular: Her üç yöntemde tek tür Candida identifikasyonunda uyumlu idi (p<0.001). Ancak birden fazla Candida türlerinin identifikasyonunda FISH ve PCR-RFLP, konvansiyonel yöntemlere göre daha duyarlı bulundu (p<0.001). Ayrıca, FISH kan kültürü pozitif örneklerden Candida türlerinin identifikasyonunda diğer iki yönteme göre daha hızlı ve ucuzdu. Birden fazla türle oluşan kandidemi oranı FISH, PCR-RFLP ve konvansiyonal yöntemle sırasıyla %20, %6 ve %4 olarak bulundu. Sonuç: Kan kültürü pozitif örneklerden Candida türlerinin hızlı identifikasyonunda PCR-RFLP ve FISH yöntemi tercih edilebilir. Ancak FISH birden fazla türle oluşan kandidemilerin belirlenmesinde daha uygun bir yöntemdir.

Anahtar Kelimeler

Candida spp.; FISH; PCR-RFLP; Candidemia; identification

Kan Kültürü Örneklerinden Candida Türlerinin Floresan In Situ Hibridizasyon (FISH), Polimeraz Zincir Reaksiyonu-Restriksiyon Fragment Uzunluk Polimorfizmi (PCR-RFLP) ve Konvansiyonel Yöntemlerle İdentifikasyonu

Öz

Amaç: Kan kültürlerinden Candida türlerinin doğru ve hızlı
identifikasyonu fungal kan dolaşımı infeksiyonlarla ilgili
mortalite ve morbiditeyi azaltmak ve etkili antifungal tedavi
sağlamak için önemlidir. Bu çalışmada, kan kültürü örneklerinden
Candida türlerinin floresan in situ hibridizasyon
(FISH), polimeraz zincir reaksiyonu-restriksiyon fragment
uzunluk polimorfizmi (PCR-RFLP) ve konvansiyonel yöntemlerle
identifikasyonu amaçlandı.
Gereçler ve Yöntemler: Candida türlerininin identifikasyonu
için 325 kan kültürü pozitif örnekten maya pozitif olan 50
örnek ile kan kültürü negatif 50 örnek FISH, PCR-RFLP ve
konvansiyonel yöntemlerle incelendi
Bulgular: Her üç yöntemde tek tür Candida identifikasyonunda
uyumlu idi (p < 0.001). Ancak birden fazla Candida
türlerinin identifikasyonunda FISH ve PCR-RFLP, konvansiyonel
yöntemlere göre daha duyarlı bulundu (p < 0.001).
Ayrıca, FISH kan kültürü pozitif örneklerden Candida türlerinin
identifikasyonunda diğer iki yönteme göre daha hızlı
ve ucuzdu. Birden fazla türle oluşan kandidemi oranı FISH,
PCR-RFLP ve konvansiyonal yöntemle sırasıyla %20, %6
ve %4 olarak bulundu.
Sonuç: Kan kültürü pozitif örneklerden Candida türlerinin
hızlı identifikasyonunda PCR-RFLP ve FISH yöntemi tercih
edilebilir. Ancak FISH birden fazla türle oluşan kandidemilerin
belirlenmesinde daha uygun bir yöntemdir.

Anahtar Kelimeler

Candida spp., FISH, PCR-RFLP, kandidemi, identifikasyon.

Kaynakça

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