Klinik açıdan önemli bazı trichophyton türlerinin hızlı tespiti için multiplex real-time pcr temelli bir yöntem geliştirilmesi
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Dosyalar
Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışma çerçevesinde, insanların deri, saç ve tırnakları gibi vücut bölgelerinde dermatolojik anlamda patojen olan Trichophyton genusuna mensup olan 6 dermatofitin geleneksel kültürasyon yöntemlerine nazaran moleküler kapsamda çok daha hızlı ve doğru tanılaması amaçlanmıştır. Trichophyton genusuna ait olan; Trichophyton interdigitale, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton tonsurans, Trichophyton verrucosum ve Trichophyton violaceum türlerinin daha önce hiçbir bilimsel çalışmada olmayan, spesifik primer ve prob oligonükleotid genetik sekanslarının in silico formatta tasarlanması amaçlanmıştır. Dermatofit küflerinin her biri için ayrı ayrı; Aktin, ATP6, Beta-Tubulin, IGS-1, ITS-1, ITS-2, ITS-3, ITS-4, Kalmodulin, NTS, RPB-1, RPB-2 ve TEF-1? gibi barkod genleri ile varsa özgün başka bir gen bölgesine yönelik çalışmalar yapılması planlanmıştır. Trichophyton interdigitale için NTS, Trichophyton mentagrophytes,
This study aims to diagnose the 6 dermatophytes belonging to the Trichophyton genus, which is dermatologically pathogen, in human body parts such as skin, hair and nails, much faster and more accurately compared to traditional culture methods in a molecular scope. It is aimed to design ever first specific primary and probe oligonucleotide genetic sequences of Trichophyton interdigitale, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton tonsurans, Trichophyton verrucosum and Trichophyton violaceum species belonging to the Trichophyton genus, in silico format. It is planned to carry out separate studies for each dermatophyte mold on barcode genes as Actin, ATP6, Beta-Tubulin, IGS-1, ITS-1, ITS-2, ITS-3, ITS-4, Calmodulin, NTS, RPB-1, RPB-2 and TEF-1? together with, if any, another unique gene region. Primer and probe sequences which are specific to target species with maximum specificity level and target NTS gene regions for Trichophyton interdigitale, VII ITS-1 gene regions for Trichophyton mentagrophytes, Trichophyton tonsurans, Trichophyton verrucosum and Trichophyton violaceum species and ITS-2 gene regions for Trichophyton rubrum are designed in silico by using various databases and with Multiplex Real-Time PCR reactions, it was determined that the design results work with full success. With Multiplex Real-Time PCR reactions, detection limits and analytical sensitivities were determined for each of the target species, it was determined that each of the reference bacterial and fungal strains and target species did not show cross reactions, and phylogenetic analyses were performed for all of the target species. Being one of the genetic methods providing much faster and more accurate diagnosis compared to traditional culture and biochemical diagnostic methods, this study will enable the clinicians to diagnose the patients with dermatophytosis infection much faster and start treatment at a much earlier time by using our species-specific designed primer and probe sets, if any target dermatophytes are available in the clinical materials taken from the concerned patients.
This study aims to diagnose the 6 dermatophytes belonging to the Trichophyton genus, which is dermatologically pathogen, in human body parts such as skin, hair and nails, much faster and more accurately compared to traditional culture methods in a molecular scope. It is aimed to design ever first specific primary and probe oligonucleotide genetic sequences of Trichophyton interdigitale, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton tonsurans, Trichophyton verrucosum and Trichophyton violaceum species belonging to the Trichophyton genus, in silico format. It is planned to carry out separate studies for each dermatophyte mold on barcode genes as Actin, ATP6, Beta-Tubulin, IGS-1, ITS-1, ITS-2, ITS-3, ITS-4, Calmodulin, NTS, RPB-1, RPB-2 and TEF-1? together with, if any, another unique gene region. Primer and probe sequences which are specific to target species with maximum specificity level and target NTS gene regions for Trichophyton interdigitale, VII ITS-1 gene regions for Trichophyton mentagrophytes, Trichophyton tonsurans, Trichophyton verrucosum and Trichophyton violaceum species and ITS-2 gene regions for Trichophyton rubrum are designed in silico by using various databases and with Multiplex Real-Time PCR reactions, it was determined that the design results work with full success. With Multiplex Real-Time PCR reactions, detection limits and analytical sensitivities were determined for each of the target species, it was determined that each of the reference bacterial and fungal strains and target species did not show cross reactions, and phylogenetic analyses were performed for all of the target species. Being one of the genetic methods providing much faster and more accurate diagnosis compared to traditional culture and biochemical diagnostic methods, this study will enable the clinicians to diagnose the patients with dermatophytosis infection much faster and start treatment at a much earlier time by using our species-specific designed primer and probe sets, if any target dermatophytes are available in the clinical materials taken from the concerned patients.
Açıklama
Anahtar Kelimeler
Trichophyton, Barkod genler, Gerçek Zamanlı PZR, Primer ve prob tasarımı, Trichophyton, Barcode genes, Real-Time PCR, Primer and probe design
