Application of High-Resolution Melting PCR to Detect the Genomic Fungal ITS 2 Region

• Published in: Laboratory medicine Vol. 51; no. 1; pp. 66 - 73
• Main Authors: Nawar, Nada N, Behiry, Iman K, Yousef, Reham H A, Emara, Mohamed A
• Format: Journal Article
• Language: English
• Published: US Oxford University Press 01.01.2020
• Subjects: Antifungal agents; Blood; Comparative analysis; Deoxyribonucleic acid; Diagnostic tests; DNA; Ethylenediaminetetraacetic acid; Fungal infections; Health aspects; Identification; Infection; Medical diagnosis; Medical examination; Medical research; Medicine, Experimental; Mortality; Mutation; Mycoses; Patients; Polymerase chain reaction; internal transcribed spacer 2; invasive fungal infections; high-resolution melting PCR; species identification; blood culture; antifungal resistance
• ISSN: 0007-5027; 1943-7730
• DOI: 10.1093/labmed/lmz034

Abstract Background Invasive fungal infections (IFIs) are a main cause of morbidity and mortality. High-resolution melting polymerase chain reaction (HRM PCR) is promising for the identification of fungal species via the detection of internal transcribed spacer 2 (ITS2). Objectives To assess the sensitivity and specificity of HRM PCR in diagnosing IFIs, compared with blood culture. Methods Our study included 100 patients who were suspected of having IFIs; we analyzed their specimens via blood culture and HRM PCR. Results Blood culture results were positive in 57 cases and negative in 43 cases. HRM PCR results were positive in 14 cases and negative in 86 cases. The 14 cases with positive results included 4 with Candida tropicalis, 4 with Candida glabrata, and 6 with Candida krusei. HRM PCR sensitivity was 24.6%, specificity was 100%, positive predictive value (PPV) was 100%, and negative predictive value (NPV) was 50%. Conclusions HRM PCR is specific but not sensitive. Blood culture is more sensitive and cannot be replaced by HRM PCR.