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Department of Myclolgy, Institute of Dermatology, Chinese Academy of Medical Scinces, Nanjing 210042, China

Objective To develop a rapid and reliable multiplex PCR procedure to detect fungi from specimens of onychomycosis.Methods Nail samples were taken from patients with clinically suspected onychomycosis.DNA was extracted from the specimens by digestion with proteinase K and boiling method.Pathogens were directly detected by triplex PCR assay based on universal fungus-,dermatophyte-,and yeast-specific primer pairs.The results were compared with those from microscopy and culture.Results One hundred and four patients were included in this study.Of them,forty-five(43.3%) were finally diagnosed as onychomycosis.For PCA,microscopy and culture,the sensitivity was 93.3%,100% and 64.4%,respectively,the speciality was 100%,86.4% and 100%,respectively,the positive predictive value was 100%,84.9% and 100%,respectively,and the negative predictive value was 95.2%,100% and 78.7%,respectively.This molecular diagnostic process needed no more than 24 h from specimen preparation to results interpretation.Conclusions The multiplex PCR assay could distinguish the three groups of mycose with a high specificity and sensitivity,shorten the detection time by approximately 2 weeks compared with culture,and provide evidence for rapid clinical diagnosis and timely mycosis treatment.