Don't throw your blood clots away: Use of blood clot may improve sensitivity of PCR diagnosis in invasive aspergillosis

E. McCulloch, G. Ramage, B. Jones, P. Warn, W. R. Kirkpatrick, Thomas F Patterson, C. Williams

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Background: The diagnosis of invasive aspergillosis (IA) remains challenging and frequently is not made until after death. Histopathological examination remains central to confirmation of diagnosis but often requires invasive procedures to obtain tissue for the examination. Detection of aspergillus DNA by quantitative PCR (qPCR) offers the potential for earlier diagnosis due to higher sensitivity, but PCR in clinical use is poorly reproducible, with different centres reporting variable results and often using different extraction and analytical methods. Aims: To optimise the performance of aspergillus PCR as a diagnostic modality. Methods: A rat inhalation model of invasive aspergillosis was used to optimise the methodology of diagnostic aspergillus PCR. Infected animals were terminally bled at 4 days post-infection; samples of EDTA blood, serum and the residual clot were pooled for subsequent analysis. DNA was extracted from each fraction using a variety of methods and an optimised qPCR reaction using an Aspergillus fumigatus primer set performed. Results: Significantly more aspergillus DNA was detected from the clot than EDTA and serum samples. Enzymatic and mechanical pretreatment reduced the yield of fungal DNA. There was some evidence that the average Ct values were greater for the EZ1 BioRobot than the MagNA Pure automated extractor, but this did not reach statistical significance at the 5% level (p = 0.078). Conclusions: Automated extraction from the clot present in a blood sample will increase DNA yield and improve the diagnostic sensitivity of the test.

Original languageEnglish (US)
Pages (from-to)539-541
Number of pages3
JournalJournal of Clinical Pathology
Volume62
Issue number6
DOIs
StatePublished - Jun 2009

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Aspergillosis
Aspergillus
Thrombosis
Polymerase Chain Reaction
DNA
Edetic Acid
Fungal DNA
Aspergillus fumigatus
Serum
Routine Diagnostic Tests
Inhalation
Early Diagnosis
Infection

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Don't throw your blood clots away : Use of blood clot may improve sensitivity of PCR diagnosis in invasive aspergillosis. / McCulloch, E.; Ramage, G.; Jones, B.; Warn, P.; Kirkpatrick, W. R.; Patterson, Thomas F; Williams, C.

In: Journal of Clinical Pathology, Vol. 62, No. 6, 06.2009, p. 539-541.

Research output: Contribution to journalArticle

McCulloch, E. ; Ramage, G. ; Jones, B. ; Warn, P. ; Kirkpatrick, W. R. ; Patterson, Thomas F ; Williams, C. / Don't throw your blood clots away : Use of blood clot may improve sensitivity of PCR diagnosis in invasive aspergillosis. In: Journal of Clinical Pathology. 2009 ; Vol. 62, No. 6. pp. 539-541.
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