PCR diagnostics of dermatophytes and other pathogenic fungi

a technology of dermatophytes and pathogenic fungi, applied in the field of pcr diagnostics of dermatophytes and other pathogenic fungi, can solve the problems of low sensitivity of techniques, false negatives, and long-term systemic treatment, and achieve the effects of reducing the number of cases, and improving the diagnostic accuracy

Inactive Publication Date: 2010-12-09
STATENS SERUM INST
View PDF2 Cites 20 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0010]The present invention solves this problem by disclosing a PCR based diagnosis of dermatophyte

Problems solved by technology

Topical therapy is sufficient in most of cases of dermatophyte skin infection, but long term and often expensive systemic treatment is necessary in cases of tinea capitis, tinea barbae and onychomycosis.
Furthermore, genus and in some cases even the species-specific diagnosis is necessary due to different susceptibility patterns and contagious potential of tha various dermatophytes, thus e.g. Microsporum species are less susceptible than Trichophyton spp. to terbinafine and should be treated with griseofulvin, T. rubrum strains are more susceptible to some antifungal drugs than T. mentagrophytes and some dermatophytes may cause epidemic outbreaks in schools and institutions due to transfer from man to man while others may not as their primary host is an animal (1
Direct microscopic examination of skin and nail material is often sufficient for the preemptive diagnosis of a fungal infection, but it does not give specif

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • PCR diagnostics of dermatophytes and other pathogenic fungi
  • PCR diagnostics of dermatophytes and other pathogenic fungi
  • PCR diagnostics of dermatophytes and other pathogenic fungi

Examples

Experimental program
Comparison scheme
Effect test

example 1

Primer Designing

[0107]The dermatophyte, Trichophyton rubrum, Microsporum canis, Trichophyton mentagrophytes-Trichophyton tonsurans complex or Epidermophyton floccosum specific primers were selected from available DNA sequences listed in Table 1. The alignment of respective sequences allowed the design of primer-pairs detecting all of the dermatophyte species (in case of panDerm1 and panDerm2 oligonucleotides), species belonging to Microsporum genus (in case of Micr773-for and Micr885-rev oligonucleotides), species belonging to Trichophyton genus (in case of Trichopyros-for and Trichopyros-rev oligonucleotides) and for specific detection of Trichophyton rubrum, Microsporum canis, Trichophyton mentagrophytes-Trichophyton tonsurans complex or Epidermophyton floccosum.

TABLE 1The sequenced used in primers designing.Gene Bank ™ accession number(www.ncbi.nih.gov)Arthoderma benhamiaeAB044155Arthoderma benhamiaeAB003558Arthoderma gypseumAB003568Arthoderma simiiAB003564Arthoderma vanbreuseghe...

example 2

[0108]Evaluation of the specificity of dermatophyte, genus Trichophyton, genus Microsporum, Trichophyton rubrum, Epidermophyton floccosum, Microsporum canis and Trichophyton mentagrophytes-Trichophyton tonsurans complex specific primers.

Materials and Methods

[0109]Strains. A list of the type, references and clinical fungal strains used in the study is presented in Table 2 Twelve strains were obtained from the National Collection of Pathogenic Fungi (United Kingdom). Clinical strains were obtained from Mycology Laboratory of Statens Serum Institute (SSI, Denmark). All clinical isolates were identified by colony characteristics and micro-morphology.

TABLE 2Microorganism used in the study.Number ofMicroorganismNCPF numberclinical isolatesMicrosporum gypseumNCPF-402Microsporum canisNCPF-17710Microsporum nanum—1Microsporum audouiniiNCPF-4365Trichophyton mentagrophytesNCPF-22410var. mentagrophytesTrichophyton mentagrophytesNCPF-780var. interdigitaleTrichophyton schoenleniniiNCPF-124—Trichop...

example 3

DNA Extraction Method—the Protocol for Invented DNA Extraction Method from Dermatophyte Infected Nails

[0125]Sixty-four nail samples were randomly chosen from samples received at the Laboratory of Mycology at SSI for microscopy and culture of dermatophytes and Candida (Table 3). Fourteen were diagnosed as T. rubrum, two as T. mentagrophytes, one as a T. tonsurans, two as Aspergillus sp., two as Candida sp., one as Alternaria sp., three as Acremonium sp. and 39 were found negative.

TABLE 3Microorganisms used in the studySpecimen diagnosed byroutine Mycology Lab asNumber of clinical isolatesTrichophyton rubrum14Trichophyton mentagrophytes2Aspergillus sp.2Candida sp.2Acremonium sp.3Alternaria1Trichophyton tonsurans1Negatives (no growth of any fungi)39

[0126]DNA from 64 clinical specimens listed in table 3 were extracted according to the following protocol:[0127]1. The nails were placed in 2 ml Eppendorf tubes[0128]2. 100 μl (can be increased up to 500) lysis buffer L1 (250 mM KCl, 60 mM N...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Temperatureaaaaaaaaaa
Temperatureaaaaaaaaaa
Temperatureaaaaaaaaaa
Login to view more

Abstract

Dermatophytes which belong to one of the three genera Epidermophyton, Trichophyton and Microsporum are the main cause of fungal infections of skin, hair and nails. Traditional diagnostic procedures consist of microscopy and culture, but due to the slow growth rate of dermatophytes typically two to four weeks are needed before a final diagnosis is obtained. The present invention is a rapid DNA extraction method extracting nucleic acids from fungi (e.g. dermatophytes and other pathogenic fungi) which can be performed from directly on hair, nail or skin specimens from humans, from naturally or experimentally infected animals or from cultured fungal colonies for the use in PCR amplification and detection assays. The present invention also includes specific primer sets for detection of any dermatophyte and for species specific detection of Trichophyton rubrum and Epidermophyton floccosum by PCR and a kit for diagnosing fungal infections.

Description

FIELD OF INVENTION[0001]A method for extracting nucleic acids from fungi, a PCR method for detecting fungi in patient samples and a PCR kit for detecting dermatophytes and diagnosing infections by the three genera Trichophyton, Microsporum and Epidermophyton. GENERAL BACKGROUND[0002]Human pathogenic dermatophytes, which belong to the three genera Trichophyton, Microsporum and Epidermophyton, are fungi that infect human skin, nails, and hair. While the genus Epidermophyton is represented only by a single species (E. floccosum), the genera Microsporum and Trichophyton include several different species (1). Depending on the site of the infection the dermatophytosis can be divided in: Tinea barbae, an infection of the bearded area mainly caused by T. verrucosum and T. mentagrophytes; Tinea capitis, usually caused by organisms from genera Microsporum and Trichophyton; Tinea corporis by any of the human pathogenic dermatophytes; Tinea favosa by T. schoenleninii; Tinea pedis frequently cau...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68C07H21/00C07H21/04
CPCC12Q1/6895C12N15/1003C12Q2600/16
Inventor BRILLOWSKA-DABROWSKA, ANNA H.
Owner STATENS SERUM INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap