Introduction of a dermatophyte polymerase chain reaction assay to the diagnostic mycology service in Scotland

C. L. Alexander, G. S. Shankland, W. Carman, C. Williams

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Dermatophytes are the major cause of superficial mycoses in samples submitted to Clinical Mycology, Glasgow. The most prevalent species is Trichophyton rubrum as identified classically by microscopy and culture. Recent advances in polymerase chain reaction (PCR) technology were examined for the feasibility of introducing a T. rubrum real-time PCR assay into a routine diagnostic service.

OBJECTIVE: To improve the diagnostic mycology service by the introduction of a real-time PCR test for T. rubrum.

METHODS: The DNA from 4972 nail and skin samples was obtained using the Qiagen QIAsymphony automated extractor. This DNA was subjected to real-time PCR using T. rubrum-specific primers and a probe.

RESULTS: During phase 1 of the study, 862 samples were analysed; 446 of 470 specimens that grew T. rubrum were detected by PCR. Out of 4110 samples analysed during phase 2, 753 T. rubrum infections were diagnosed and reported within 72 h. A total of 3357 samples were negative for a fungal infection by PCR and microscopy; these were also reported within 72 h.

CONCLUSIONS: A vast reduction in the turnaround times can be achieved using this technique as opposed to classical methods. Samples which are PCR negative but microscopy positive are still subjected to culture. Screening samples for their suitability for PCR prior to processing eliminates the application of PCR for T. rubrum on inappropriate samples such those from the scalp or pityriasis versicolor.

Original languageEnglish
Pages (from-to)966-972
Number of pages7
JournalThe British Journal of Dermatology
Volume164
Issue number5
DOIs
Publication statusPublished - May 2011
Externally publishedYes

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Mycology
Diagnostic Services
Arthrodermataceae
Scotland
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Microscopy
Mycoses
Tinea Versicolor
Trichophyton
DNA
Nails
Scalp
Technology
Skin
Infection

Keywords

  • DNA, Fungal
  • Dermatomycoses
  • Humans
  • Mass Screening
  • Mycology
  • Nails
  • Polymerase Chain Reaction
  • Scotland
  • Sensitivity and Specificity
  • Skin
  • Trichophyton
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

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title = "Introduction of a dermatophyte polymerase chain reaction assay to the diagnostic mycology service in Scotland",
abstract = "BACKGROUND: Dermatophytes are the major cause of superficial mycoses in samples submitted to Clinical Mycology, Glasgow. The most prevalent species is Trichophyton rubrum as identified classically by microscopy and culture. Recent advances in polymerase chain reaction (PCR) technology were examined for the feasibility of introducing a T. rubrum real-time PCR assay into a routine diagnostic service.OBJECTIVE: To improve the diagnostic mycology service by the introduction of a real-time PCR test for T. rubrum.METHODS: The DNA from 4972 nail and skin samples was obtained using the Qiagen QIAsymphony automated extractor. This DNA was subjected to real-time PCR using T. rubrum-specific primers and a probe.RESULTS: During phase 1 of the study, 862 samples were analysed; 446 of 470 specimens that grew T. rubrum were detected by PCR. Out of 4110 samples analysed during phase 2, 753 T. rubrum infections were diagnosed and reported within 72 h. A total of 3357 samples were negative for a fungal infection by PCR and microscopy; these were also reported within 72 h.CONCLUSIONS: A vast reduction in the turnaround times can be achieved using this technique as opposed to classical methods. Samples which are PCR negative but microscopy positive are still subjected to culture. Screening samples for their suitability for PCR prior to processing eliminates the application of PCR for T. rubrum on inappropriate samples such those from the scalp or pityriasis versicolor.",
keywords = "DNA, Fungal, Dermatomycoses, Humans, Mass Screening, Mycology, Nails, Polymerase Chain Reaction, Scotland, Sensitivity and Specificity, Skin, Trichophyton, Journal Article, Research Support, Non-U.S. Gov't",
author = "Alexander, {C. L.} and Shankland, {G. S.} and W. Carman and C. Williams",
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Introduction of a dermatophyte polymerase chain reaction assay to the diagnostic mycology service in Scotland. / Alexander, C. L.; Shankland, G. S.; Carman, W.; Williams, C.

In: The British Journal of Dermatology, Vol. 164, No. 5, 05.2011, p. 966-972.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Introduction of a dermatophyte polymerase chain reaction assay to the diagnostic mycology service in Scotland

AU - Alexander, C. L.

AU - Shankland, G. S.

AU - Carman, W.

AU - Williams, C.

N1 - © 2011 The Authors. BJD © 2011 British Association of Dermatologists.

PY - 2011/5

Y1 - 2011/5

N2 - BACKGROUND: Dermatophytes are the major cause of superficial mycoses in samples submitted to Clinical Mycology, Glasgow. The most prevalent species is Trichophyton rubrum as identified classically by microscopy and culture. Recent advances in polymerase chain reaction (PCR) technology were examined for the feasibility of introducing a T. rubrum real-time PCR assay into a routine diagnostic service.OBJECTIVE: To improve the diagnostic mycology service by the introduction of a real-time PCR test for T. rubrum.METHODS: The DNA from 4972 nail and skin samples was obtained using the Qiagen QIAsymphony automated extractor. This DNA was subjected to real-time PCR using T. rubrum-specific primers and a probe.RESULTS: During phase 1 of the study, 862 samples were analysed; 446 of 470 specimens that grew T. rubrum were detected by PCR. Out of 4110 samples analysed during phase 2, 753 T. rubrum infections were diagnosed and reported within 72 h. A total of 3357 samples were negative for a fungal infection by PCR and microscopy; these were also reported within 72 h.CONCLUSIONS: A vast reduction in the turnaround times can be achieved using this technique as opposed to classical methods. Samples which are PCR negative but microscopy positive are still subjected to culture. Screening samples for their suitability for PCR prior to processing eliminates the application of PCR for T. rubrum on inappropriate samples such those from the scalp or pityriasis versicolor.

AB - BACKGROUND: Dermatophytes are the major cause of superficial mycoses in samples submitted to Clinical Mycology, Glasgow. The most prevalent species is Trichophyton rubrum as identified classically by microscopy and culture. Recent advances in polymerase chain reaction (PCR) technology were examined for the feasibility of introducing a T. rubrum real-time PCR assay into a routine diagnostic service.OBJECTIVE: To improve the diagnostic mycology service by the introduction of a real-time PCR test for T. rubrum.METHODS: The DNA from 4972 nail and skin samples was obtained using the Qiagen QIAsymphony automated extractor. This DNA was subjected to real-time PCR using T. rubrum-specific primers and a probe.RESULTS: During phase 1 of the study, 862 samples were analysed; 446 of 470 specimens that grew T. rubrum were detected by PCR. Out of 4110 samples analysed during phase 2, 753 T. rubrum infections were diagnosed and reported within 72 h. A total of 3357 samples were negative for a fungal infection by PCR and microscopy; these were also reported within 72 h.CONCLUSIONS: A vast reduction in the turnaround times can be achieved using this technique as opposed to classical methods. Samples which are PCR negative but microscopy positive are still subjected to culture. Screening samples for their suitability for PCR prior to processing eliminates the application of PCR for T. rubrum on inappropriate samples such those from the scalp or pityriasis versicolor.

KW - DNA, Fungal

KW - Dermatomycoses

KW - Humans

KW - Mass Screening

KW - Mycology

KW - Nails

KW - Polymerase Chain Reaction

KW - Scotland

KW - Sensitivity and Specificity

KW - Skin

KW - Trichophyton

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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M3 - Article

VL - 164

SP - 966

EP - 972

JO - The British Journal of Dermatology

JF - The British Journal of Dermatology

SN - 0007-0963

IS - 5

ER -