Diagnostic value of a nested polymerase chain reaction for diagnosing cutaneous sporotrichosis from paraffin‐embedded skin tissue

• Published in: Mycoses Vol. 62; no. 12; pp. 1148 - 1153
• Main Authors: Hayashi, Shujiro, Kaminaga, Tomoko, Baba, Akiko, Koike, Saiko, Koike, Masami, Kanno, Miki, Ishikawa, Satoko, Tsukada, Kyoujyu, Suzuki, Toshihiro, Hamasaki, Yoichiro, Sairenchi, Toshimi, Kobashi, Gen, Igawa, Ken
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01-12-2019
• Subjects: Aged; Anesthesia; Biopsy; Diagnosis; DNA, Fungal - isolation & purification; Female; Fungi; Humans; Male; Middle Aged; Molecular Diagnostic Techniques - standards; Paraffin; Paraffin Embedding; PCR; Polymerase chain reaction; Polymerase Chain Reaction - standards; Sensitivity and Specificity; Skin - microbiology; skin infection; Sporothrix; Sporothrix - isolation & purification; Sporothrix schenckii; Sporotrichosis; Sporotrichosis - diagnosis; Sporothrix schenckii; sporotrichosis; skin infection; PCR
• ISSN: 0933-7407; 1439-0507
• DOI: 10.1111/myc.13004

Summary
Background
The gold standard for diagnosis of cutaneous sporotrichosis involves the isolation of the fungus, Sporothrix, by a culture test. Generally, the sampling for the culture test is performed at the same time as skin biopsy under local anaesthesia. However, the culture test may occasionally return a false negative result.
Objective
The aim of our study was to investigate the diagnostic value of a molecular method for diagnosing cutaneous sporotrichosis from formalin‐fixed and paraffin‐embedded (FFPE) tissues.
Methods
Over a 30‐year period, we collected 52 cases of cutaneous sporotrichosis from biopsied specimens that had been positively diagnosed by a culture test. A nested PCR specific for Sporothrix detection was applied using FFPE tissue as template. The results were compared with control samples from 79 patients diagnosed with other cutaneous diseases according to histopathological, clinical findings and a cutler test.
Results
Of the 52 patients who were tested positive on the culture test, all cutaneous diseases were detected by PCR. Of the 59 patients in the control group, 58 tested negative by PCR. Under our conditions, the calculated sensitivity of this method was 100%, the specificity was 98.7% and the kappa coefficient was 0.984 (95% CI: 0.953‐1.000).
Conclusions
The specific PCR assay used appears to be a useful tool for the prompt and accurate diagnosis of sporotrichosis. Using this method, it would be possible to diagnose cutaneous sporotrichosis for patients who were suspected of cutaneous sporotrichosis but tested negative on culturing, and for pathologically suspected cutaneous sporotrichosis patients for whom the culture test was not undertaken.