Abstract
Amaç: Bu çalışmanın amacı basit bir besiyeri olan Mueller-Hinton agar (MHA)’ın germ tüp testinde (GTT)
kullanılabilirliğinin araştırılmasıdır.
Yöntem: Çeşitli klinik örneklerden enfeksiyon etkeni olarak izole edilen toplam 86 Candida suşu çalışmaya alındı.
Tüm izolatlar Sabouraud Dekstroz Agar (SDA) (Salubris, Türkiye)’a pasajlanarak, 37°C’de 24-48 saat inkübe
edildi. İzolatlar VITEK-MS (Biomérieux, Fransa) cihazı ile tanımlandı. Serum GTT için tek bir Candida kolonisi 0.5
ml taze insan serumuna inoküle edildi. MHA GTT için tek koloniden MHA yüzeyine Dalmau tekniği ile çizgi ekimi
yapıldı ve üzeri steril lamel ile kapatıldı. 37°C’de 1.5 ve 3 saat inkübasyon sonrasında tüm serumlar ve MHA plakları
ışık mikroskobu ile değerlendirildi. Duyarlılık ve özgüllük değerlendirmesinde serum GTT altın standart yöntem
olarak kabul edildi.
Bulgular: Tanımlama sonuçları şöyleydi: 51 Candida albicans, bir C. dubliniensis, 12 C. parapsilosis, 12 C. glabrata, 4 C.
kefyr, 3 C. tropicalis, 3 C. krusei. Serum GTT’nde 1.5 saat sonunda 30 (%58.8), 3 saat sonunda ise 36 (%70.6) C.
albicans izolatı pozitif olarak bulundu. MHA GTT’nde ise 1.5 saat sonunda 21 (%41.2), 3 saat sonunda ise 40
(%78.4) C. albicans izolatı pozitif olarak saptandı. C. dubliniensis dışındaki tüm non-albicans türleri her iki testte de
negatif olarak bulundu. C. dubliniensis her iki testte de 1.5 ve 3 saatte GTT pozitif olarak saptandı. MHA’ın 1.5
saatte duyarlılığı %58, özgüllüğü %92; 3 saatte ise duyarlılığı %86, özgüllüğü %81 olarak hesaplandı.
Sonuç: MHA, GTT için güvenli ve kolay bulunabilen bir besiyeridir ve serum GTT’ne alternatif olarak kullanılabilir.
MHA’da GTT araştırılması esnasında, daha yüksek duyarlılığa sahip olduğundan dolayı, 1.5 saat yerine 3 saatlik
inkübasyon önerilmektedir.
References
- Hazen KC, Howell SA. Candida, Cryptococcus, and Other Yeasts of Medical Importance. Manual of Clinical Microbiology, 9.ed. Murray PR, Baron EJ, Jorgensen JH, Landry ML, Pfaller MA (ed), Washington, DC. ASM Press, 2007; s. 1762-1788
- Edwards JE, Candida Species. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases. 7. ed. Mandell GL, Bennett JE, Dolın R. (ed), Philadelphia, PA. Churchill Livingstone, 2010; s. 3225 - 3240.
- Sudbery P, Gow N, Berman J. The distinct morphogenic states of Candida albicans. Trends in Microbiol, 2004; 12(7): 317-324. Mackenzie DWR. Serum tube identification of Candida albicans. Journ Clin Pathol, 1962; 15(6): 563-565.
- Sullivan D, Coleman D. Candida dubliniensis: characteristics and identification. Journ Clin Microbiol, 1998; 36(2): 329-334.
- Beheshti F, Smith AG, Krause GW. Germ tube and chlamydospore formation by Candida albicans on a new medium. Journ Clin Microbiol, 1975; 2(4): 345-348.
- Warwood NM, Blazevic DJ. Comparison of cream of rice agar and horse serum for differentiating germ tubes of Candida albicans from filaments of Candida tropicalis. Journ Clin Microbiol, 1977; 5(4): 501-502.
- Yong DC, Smitka C, Prytula A ve diğ. The comparison of two agar media for germ tube and chlamydospore production by Candida albicans. Health lab Sci, 1978; 15(4): 197-200.
- Rimek D, Fehse B, Göpel P. Evaluation of Mueller‐Hinton‐agar as a simple medium for the germ tube production of Candida albicans and Candida dubliniensis. Mycoses, 2008; 51(3): 205-208.
- Mattei AS, Alves SH, Severo CB ve diğ. Use Of Mueller-Hınton Broth And Agar In The Germ Tube Test. Revista do Instituto de Medicina Tropical de São Paulo, 2014; 56(6): 483-485.
- Hoppe JE, Frey P. Evaluation of six commercial tests and the germ-tube test for presumptive identification of Candida albicans. Eur J Clin Microbiol Infect Dis, 1999; 18(3): 188-191.
- Arora DR, Saini S, Gupta N. Evaluation of germ tube test in various media. Indian J Pathol & Microbiol, 2003; 46(1): 124-126.
- Hilmioglu S, Ilkit M, Badak Z. Comparison of 12 liquid media for germ tube production of Candida albicans and C. tropicalis. Mycoses, 2007; 50(4): 282-285.
- Atalay A, Koç AN, Parkan Ö ve diğ. Çimlenme borusu (germ tüp) testinde Mueller-Hinton Agar serumların yerini alabilir mi? 1. Ulusal Tıbbi Mikoloji Kongresi, Poster no: 07, 25 Eylül 2014, Ankara
Abstract
Aims: The aim of this study is to evaluate Mueller-Hinton Agar (MHA) as a simple medium for the determination of
germ tube test (GTT).
Methods: A total of 86 Candida species isolated from different clinical specimens were included. All isolates were
cultured onto Sabouraud Dextrose Agar (Salubris, Turkey) and incubated at 37°C for 24-48 hours. They were
identified by VITEK MS (Biomérieux, France). For serum GTT, a single Candida colony was inoculated into 0.5 ml
freshly obtained human serum. For MHA GTT, an inoculum of single colony was streaked by Dalmau technique onto
MHA and covered by a sterile coverslip. After incubation at 37°C for 1.5 h and 3 h, all sera and MHA plates were
examined using a light microscope. Serum GTT was accepted as gold standard method for the determination of
sensitivity and specificity values.
Results: The identification results were as follows: 51 Candida albicans, one C. dubliniensis, 12 C. parapsilosis, 12
C. glabrata, 4 C. kefyr, 3 C. tropicalis, 3 C. krusei. Serum GTT was positive in 30 (58.8%) and 36 (70.6%) of C.
albicans isolates at 1.5 h and 3 h, respectively. MHA GTT was positive in 21 (41.2%) and 40 (78.4%) of C. albicans
isolates at 1.5 h and 3 h, respectively. All non-albicans species except C. dubliniensis were found as negative using
both tests. Both GTTs were positive for C. dubliniensis at 1.5 h and 3 h. MHA sensitivity and specificity was
determined as 58% and 92% at 1.5 h; 86% and 81% at 3 h, respectively.
Conclusions: MHA is safer and easier medium for GTT that may be used as alternative to serum GTT. The
evaluation of GTT on MHA after 3 h incubation is recommended since it has higher sensitivity compared to 1.5 h.
References
- Hazen KC, Howell SA. Candida, Cryptococcus, and Other Yeasts of Medical Importance. Manual of Clinical Microbiology, 9.ed. Murray PR, Baron EJ, Jorgensen JH, Landry ML, Pfaller MA (ed), Washington, DC. ASM Press, 2007; s. 1762-1788
- Edwards JE, Candida Species. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases. 7. ed. Mandell GL, Bennett JE, Dolın R. (ed), Philadelphia, PA. Churchill Livingstone, 2010; s. 3225 - 3240.
- Sudbery P, Gow N, Berman J. The distinct morphogenic states of Candida albicans. Trends in Microbiol, 2004; 12(7): 317-324. Mackenzie DWR. Serum tube identification of Candida albicans. Journ Clin Pathol, 1962; 15(6): 563-565.
- Sullivan D, Coleman D. Candida dubliniensis: characteristics and identification. Journ Clin Microbiol, 1998; 36(2): 329-334.
- Beheshti F, Smith AG, Krause GW. Germ tube and chlamydospore formation by Candida albicans on a new medium. Journ Clin Microbiol, 1975; 2(4): 345-348.
- Warwood NM, Blazevic DJ. Comparison of cream of rice agar and horse serum for differentiating germ tubes of Candida albicans from filaments of Candida tropicalis. Journ Clin Microbiol, 1977; 5(4): 501-502.
- Yong DC, Smitka C, Prytula A ve diğ. The comparison of two agar media for germ tube and chlamydospore production by Candida albicans. Health lab Sci, 1978; 15(4): 197-200.
- Rimek D, Fehse B, Göpel P. Evaluation of Mueller‐Hinton‐agar as a simple medium for the germ tube production of Candida albicans and Candida dubliniensis. Mycoses, 2008; 51(3): 205-208.
- Mattei AS, Alves SH, Severo CB ve diğ. Use Of Mueller-Hınton Broth And Agar In The Germ Tube Test. Revista do Instituto de Medicina Tropical de São Paulo, 2014; 56(6): 483-485.
- Hoppe JE, Frey P. Evaluation of six commercial tests and the germ-tube test for presumptive identification of Candida albicans. Eur J Clin Microbiol Infect Dis, 1999; 18(3): 188-191.
- Arora DR, Saini S, Gupta N. Evaluation of germ tube test in various media. Indian J Pathol & Microbiol, 2003; 46(1): 124-126.
- Hilmioglu S, Ilkit M, Badak Z. Comparison of 12 liquid media for germ tube production of Candida albicans and C. tropicalis. Mycoses, 2007; 50(4): 282-285.
- Atalay A, Koç AN, Parkan Ö ve diğ. Çimlenme borusu (germ tüp) testinde Mueller-Hinton Agar serumların yerini alabilir mi? 1. Ulusal Tıbbi Mikoloji Kongresi, Poster no: 07, 25 Eylül 2014, Ankara
Details
| Subjects | Health Care Administration |
|---|---|
| Journal Section | Original Article |
| Authors | |
| Publication Date | September 30, 2015 |
| Submission Date | November 6, 2017 |
| Acceptance Date | September 15, 2015 |
| Published in Issue | Year 2015 Volume: 1 Issue: 1 |
