Rapid detection of dermatophytes and Candida albicans in onychomycosis specimens by an oligonucleotide array

BMC Infectious Diseases

Tsung Chain Chang¹  Hsien Chang Chang²  Hsin Chieh Li¹  Hsin Yi Hsieh¹  Chao-Kai Hsu³  Jong Soo Choi⁵  Mark Ming-Long Hsu⁴  Huan Wen Han² 

[1] Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan, Taiwan;Institute of Biomedical Engineering, College of Engineering, National Cheng Kung University, Tainan, Taiwan;Department of Dermatology, National Cheng Kung University Hospital, Tainan, Taiwan;ESkin Clinic, Tainan, Taiwan;Department of Dermatology, Yeungnam University College of Medicine, Daegu, Korea

关键词: Dermatophytes;    Internal transcribed spacer;    Molecular diagnosis;    Oligonucleotide array;    Onychomycosis;   

Others  :  1125391
DOI  :  10.1186/s12879-014-0581-5

 received in 2013-09-02, accepted in 2014-10-22,  发布年份 2014

【 摘 要 】

Background

Onychomycosis is a fungal infection of nails, leading to the gradual destruction of the nail plate. Treatment of onychomycosis may need long-time oral antifungal therapy that can have potential side effects, thus accurate diagnosis of the disease before treatment is important. Culture for diagnosis of onychomycosis is time-consuming and has high false-negative rates. To expedite the diagnosis, an oligonucleotide array, based on hybridization between immobilized oligonucleotide probes and PCR products, for direct detection of dermatophytes and Candida albicans in clinical specimens was evaluated.

Methods

Species-specific oligonucleotide probes designed from the internal transcribed spacer (ITS) regions of the rRNA gene were immobilized on a nylon membrane. The assay procedures consisted of PCR amplification of the ITS using universal primers, followed by hybridization of the digoxigenin-labeled amplicons to probes on the array. Thirty two nail samples (29 patients) were analyzed by the array, and the results were compared with those obtained by culture. Array-positive but culture-negative samples were confirmed by cloning and re-sequencing of the amplified ITS and by reviewing patient’s clinical data. The total recovery of culture and confirmed array-positive but culture-negative results was considered 100% and was used for performance evaluation of both methods.

Results

Concordant results were obtained in 21 samples (10 positives and 11 negatives) by both methods. Eleven samples were array-positive but culture-negative; among them, 9 samples were considered true positives after discrepant analysis. Comparing with culture, the array had significantly higher sensitivity [100% (95% CI 82.2% −100%) vs 52.6% (28.9% −75.5%), p <0.001] and negative predictive value [100% (71.3% −100%) vs 59.1% (36.4% −79.3%), p <0.05), while no significant differences were observed in specificity (84.6% vs 100%, p =0.48) and positive predictive value (90.5% vs 100%, p =1.0). The whole procedures of the array were about 24 h, whilst results from culture take 1 to 3 weeks.

Conclusions

The array offers an accurate and rapid alternative to culture. Rapid diagnosis can expedite appropriate antifungal treatment of onychomycosis. However, the single site nature of this study conducted at a referral hospital invites caution.

【 授权许可】

   
2014 Han et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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