Real-time PCR method for detection of zygomycetes

D. Jane Hata, Seanne P. Buckwalter, Bobbi S. Pritt, Glenn D. Roberts, Nancy L. Wengenack

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

Zygomycete infections can be devastating in immunocompromised hosts. Difficulties in the histopathologic differentiation of this class from other filamentous fungi (e.g., Aspergillus spp., Fusarium spp.) may lead to delays in diagnosis and initiation of appropriate treatment, thereby significantly affecting patient outcome. A real-time PCR assay was developed to detect species of the zygomycete genera Absidia, Apophysomyces, Cunninghamella, Mucor, Rhizopus, and Saksenaea in culture and tissue samples. Primers and fluorescence resonance energy transfer hybridization probes were designed to detect a 167-bp conserved region of the multicopy zygomycete cytochrome b gene. A plasmid containing target sequence from Mucor racemosus was constructed as a positive control. The analytical sensitivity of the assay is 10 targets/μl, and a specificity panel consisting of other filamentous fungi, yeasts (Candida spp.), and bacteria demonstrated no cross-reactivity in the assay. The clinical sensitivity and specificity of the assay from culture isolates were 100% (39/39) and 92% (59/64), respectively. Sensitivity and specificity determined using a limited number of fresh tissue specimens were both 100% (2/2). The sensitivity seen with formalin-fixed, paraffin-embedded tissues was 56% (35/62), and the specificity was 100% (19/19). The speed, sensitivity, and specificity of the PCR assay indicate that it is useful for the rapid and accurate detection of zygomycetes.

Original languageEnglish (US)
Pages (from-to)2353-2358
Number of pages6
JournalJournal of Clinical Microbiology
Volume46
Issue number7
DOIs
StatePublished - Jul 2008

Fingerprint

Mucor
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Fungi
Absidia
Cunninghamella
Rhizopus
Fluorescence Resonance Energy Transfer
Cytochromes b
Fusarium
Immunocompromised Host
Aspergillus
Candida
Paraffin
Formaldehyde
Plasmids
Yeasts
Bacteria
Polymerase Chain Reaction
Infection

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Hata, D. J., Buckwalter, S. P., Pritt, B. S., Roberts, G. D., & Wengenack, N. L. (2008). Real-time PCR method for detection of zygomycetes. Journal of Clinical Microbiology, 46(7), 2353-2358. https://doi.org/10.1128/JCM.02331-07

Real-time PCR method for detection of zygomycetes. / Hata, D. Jane; Buckwalter, Seanne P.; Pritt, Bobbi S.; Roberts, Glenn D.; Wengenack, Nancy L.

In: Journal of Clinical Microbiology, Vol. 46, No. 7, 07.2008, p. 2353-2358.

Research output: Contribution to journalArticle

Hata, DJ, Buckwalter, SP, Pritt, BS, Roberts, GD & Wengenack, NL 2008, 'Real-time PCR method for detection of zygomycetes', Journal of Clinical Microbiology, vol. 46, no. 7, pp. 2353-2358. https://doi.org/10.1128/JCM.02331-07
Hata DJ, Buckwalter SP, Pritt BS, Roberts GD, Wengenack NL. Real-time PCR method for detection of zygomycetes. Journal of Clinical Microbiology. 2008 Jul;46(7):2353-2358. https://doi.org/10.1128/JCM.02331-07
Hata, D. Jane ; Buckwalter, Seanne P. ; Pritt, Bobbi S. ; Roberts, Glenn D. ; Wengenack, Nancy L. / Real-time PCR method for detection of zygomycetes. In: Journal of Clinical Microbiology. 2008 ; Vol. 46, No. 7. pp. 2353-2358.
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