Journal of the European Academy of Dermatology and Venereology : JEADV · 2018
Comparison of fungal fluorescent staining and ITS rDNA PCR-based sequencing with conventional methods for the diagnosis of onychomycosis.
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Bao F, Fan Y, Sun L, Yu Y, Wang Z, Pan Q, Yu C, Liu H, Zhang F
Аннотация
BACKGROUND: The current gold standard for diagnosing onychomycosis is direct microscopic examination and culturing. Fungal culture is a time-consuming procedure, while direct microscopy of potassium hydroxide (KOH) mounts suffers from low sensitivity. More rapid and sensitive methods for the diagnosis of onychomycosis are in high demand.
OBJECTIVE: To establish an effective method for the diagnosis of onychomycosis by assessing the efficacies of fungal fluorescent staining and internal transcribed spacer (ITS) ribosomal DNA (rDNA) polymerase chain reaction (PCR)-based sequencing.
METHODS: A total of 204 clinical specimens from patients with suspected onychomycosis were analysed. The gold standard for a true positive sample was positive by KOH, culturing or both methods. All specimens were also tested by fungal fluorescent staining and ITS rDNA PCR-based sequencing. We compared the detection, sensitivity and specificity for these two methods with conventional methods.
RESULTS: In total, 126 (62%) and 102 (50%) were detected by fluorescent staining and PCR-based sequencing, respectively. According to the conventional diagnostic standard, the sensitivity of fluorescent staining and PCR-based sequencing was 97% and 78%, respectively, and specificities of 89% and 90%, respectively. Use of fluorescence enhanced the sensitivity of direct examination by 12% compared with KOH. PCR-based sequencing increased the sensitivity by 6% compared with culturing.
CONCLUSIONS: Fluorescence microscopy has a higher sensitivity for the detection of fungi in nail specimens compared with KOH and can be used as a rapid screening tool. PCR-based sequencing was faster and more sensitive compared with culture and when used in conjunction with fluorescence microscopy resulted in higher efficiency.
OBJECTIVE: To establish an effective method for the diagnosis of onychomycosis by assessing the efficacies of fungal fluorescent staining and internal transcribed spacer (ITS) ribosomal DNA (rDNA) polymerase chain reaction (PCR)-based sequencing.
METHODS: A total of 204 clinical specimens from patients with suspected onychomycosis were analysed. The gold standard for a true positive sample was positive by KOH, culturing or both methods. All specimens were also tested by fungal fluorescent staining and ITS rDNA PCR-based sequencing. We compared the detection, sensitivity and specificity for these two methods with conventional methods.
RESULTS: In total, 126 (62%) and 102 (50%) were detected by fluorescent staining and PCR-based sequencing, respectively. According to the conventional diagnostic standard, the sensitivity of fluorescent staining and PCR-based sequencing was 97% and 78%, respectively, and specificities of 89% and 90%, respectively. Use of fluorescence enhanced the sensitivity of direct examination by 12% compared with KOH. PCR-based sequencing increased the sensitivity by 6% compared with culturing.
CONCLUSIONS: Fluorescence microscopy has a higher sensitivity for the detection of fungi in nail specimens compared with KOH and can be used as a rapid screening tool. PCR-based sequencing was faster and more sensitive compared with culture and when used in conjunction with fluorescence microscopy resulted in higher efficiency.