PCR-reverse blot hybridization assay for fast and accurate identification of causative species in superficial fungal infections

Abstract

Compared with KOH preparation or fungal culture, PCR-REBA has higher sensitivity and specificity. Therefore, PCR-REBA could be a useful method in clinical settings because it can identify species quickly and accurately, and can also determine the existence of pathogens.

Simultaneous detection of barrier- and immune-related gene variations in patients with atopic dermatitis by reverse blot hybridization assay

“…This study is also significant in that we simultaneously assessed a total of 35 gene variants using REBA with only a small amount of tissue sample. REBA, along with DNA chip assays, has been used not only to identify genotypes of human papilloma virus, mycobacteria and dermatophytes, but also to identify mutations within specific genes . This trial is significant because it is the first application of REBA to identify human disease‐related gene variations, especially in patients with AD with complex gene variations.…”

“…The REBA membrane was designed to contain a total of 70 probes [35 wild‐type (WT) and 35 mutant‐type (MT) probes] to detect these genes, and one control probe for colorimetric signal detection (Table ). REBA was performed in accordance with our previous reports (Fig. ).…”

“…To design specific primer molecules, the sequences of KLK7 , SPINK5 , FLG , DEFB1 , TNF α, KDR , FCER1A, IL4, IL5, IL5RA, IL9, IL10, IL12, IL12R, IL13 and IL18 were compiled from the GenBank database of the National Center for Biotechnology Information (NCBI). Multiplex PCR was performed in accordance with our previous reports . The sequences of the PCR products were analysed.…”

“…Multiplex PCR was performed in accordance with our previous reports. 19,20 The sequences of the PCR products were analysed. Direct sequencing of both strands of the PCR product was carried out on an ABI 3100 Genetic Analyzer) by COSMO Genetech Co. Ltd. (Seoul, Korea) to verify the accuracy of REBA.…”

Prevalence and Characteristics of Onychomycosis in Patients with Knee Osteoarthritis: A Single-centre Prospective Cross-sectional Study

“…PCR-based REBA is a molecular biologic method that can be used for diagnosing bacterial or fungal species ( 37 – 40 ). Studies showed that REBA yields fast and reliable results with higher specificity and sensitivity compared with fungal culture or KOH test ( 38 , 40 ). The sensitivity and NPV of the KOH test relative to REBA were only moderate, at 66.2% and 65.7%, respectively.…”