Identification of Phytophthora Species by a High Resolution Melting Analysis: an Innovative Tool for Rapid Differentiation
نویسندگان
چکیده
Zambounis A., Samaras A., Xanthopoulou A., Osathanunkul M., Schena L., Tsaftaris A., Madesis P. (2016): Identification of Phytophthora species by a high resolution melting analysis: an innovative tool for rapid differentiation. Plant Protect. Sci., 52. A new molecular method via the high resolution melting (HRM) analysis of the Ypt1 gene non-coding regions was validated for ten Phytophthora species with a broad host range from forest trees to crop species. The melting curve analysis of the amplicons specifically grouped all species into 10 respective unique and distinct HRM curve profiles. The analysis of the normalised HRM melting curves, assigning P. nicotianae as a normalised reference genotype, revealed that the genotype similarities among all the species were adequately low, indicating that Ypt1 marker was sufficient to identify and differentiate the tested species. This HRM method is rapid and reproducible allowing the identification of Phytophthora species and the screening of eventual variants eliminating the separate steps and reducing the risk of contamination.
منابع مشابه
Rapid identification of Phytophthora ramorum using PCR-SSCP analysis of ribosomal DNA ITS-1.
AIMS The primary objectives of this study were to determine if a single-strand conformation polymorphism (SSCP) analysis can be used for rapid identification of Phytophthora ramorum, an important quarantine plant pathogen worldwide, and to further assess the potential of the SSCP technique as a taxonomic tool for the genus Phytophthora. METHODS AND RESULTS SSCP of ribosomal DNA internal trans...
متن کاملDevelopment of a High-Resolution Melting Method for Screening R188H Polymorphism in XRCC2 Gene
Background: The High Resolution Melting (HRM) method is a new scanning method for detecting unknown changes in DNA and its advantages have persuaded researchers to recruit it as a screening method. Objectives: Here, we developed a HRM method to screen R188H SNP (rs3218536) of XRCC2 and compared the results with a well known PCR-RFLP technique. Materials and Methods: Genomic ...
متن کاملRapid detection and identification of clinically important bacteria by high-resolution melting analysis after broad-range ribosomal RNA real-time PCR.
BACKGROUND Broad-range PCR provides valuable information for detecting bacterial infections. This study assesses the combined use of broad-range real-time PCR and high-resolution melting analysis for rapid detection and identification of clinically important bacteria. METHODS We subjected 46 bacterial culture colonies representing 25 clinically important bacterial species to LightCycler real-...
متن کاملMolecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis
Human schistosomiasis caused by Schistosoma japonicum and Schistosoma mekongi is a chronic and debilitating helminthic disease still prevalent in several countries of Asia. Due to morphological similarities of cercariae and eggs of these 2 species, microscopic differentiation is difficult. High resolution melting (HRM) real-time PCR is developed as an alternative tool for the detection and diff...
متن کاملPCR-restriction enzyme method based on the polymorphism in ribosomal-DNA for identification of the most important dermatophyte species in Iran
Background and aim: Dermatophytosis (tinea, ringworm) is the infection of skin, hair or nail that is caused by various keratinophilic fungi (dermatophytes). Dermatophytosis is a common infection throughout the world including all parts of Iran. As conventional laboratory procedures for identification of different dermatophytes are slow or lack specificity, more rapid and reliable methods are st...
متن کامل