Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples

Bertie H C G M de Leeuw; W Sebastiaan Voskuil; Boulos Maraha; Anneke van der Zee; Pieter J Westenend; Johannes G Kusters

doi:10.1016/j.yexmp.2015.03.021

Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples

Bertie H C G M de Leeuw, W Sebastiaan Voskuil, Boulos Maraha, Anneke van der Zee, Pieter J Westenend, Johannes G Kusters

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection rates. In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples.

Original language	English
Pages (from-to)	390-392
Number of pages	3
Journal	Experimental and molecular pathology
Volume	98
Issue number	3
DOIs	https://doi.org/10.1016/j.yexmp.2015.03.021
Publication status	Published - Jun 2015

Keywords

Bronchoalveolar Lavage Fluid
DNA, Bacterial
Formaldehyde
Humans
Paraffin Embedding
Pneumocystis jirovecii
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Tissue Fixation

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10.1016/j.yexmp.2015.03.021

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title = "Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples",

abstract = "The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection rates. In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples.",

keywords = "Bronchoalveolar Lavage Fluid, DNA, Bacterial, Formaldehyde, Humans, Paraffin Embedding, Pneumocystis jirovecii, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Tissue Fixation",

author = "{de Leeuw}, {Bertie H C G M} and Voskuil, {W Sebastiaan} and Boulos Maraha and {van der Zee}, Anneke and Westenend, {Pieter J} and Kusters, {Johannes G}",

year = "2015",

month = jun,

doi = "10.1016/j.yexmp.2015.03.021",

language = "English",

volume = "98",

pages = "390--392",

journal = "Experimental and molecular pathology",

issn = "0014-4800",

publisher = "Academic Press Inc.",

number = "3",

}

Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples. / de Leeuw, Bertie H C G M; Voskuil, W Sebastiaan; Maraha, Boulos et al.
In: Experimental and molecular pathology, Vol. 98, No. 3, 06.2015, p. 390-392.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples

AU - de Leeuw, Bertie H C G M

AU - Voskuil, W Sebastiaan

AU - Maraha, Boulos

AU - van der Zee, Anneke

AU - Westenend, Pieter J

AU - Kusters, Johannes G

PY - 2015/6

Y1 - 2015/6

N2 - The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection rates. In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples.

AB - The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection rates. In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples.

KW - Bronchoalveolar Lavage Fluid

KW - DNA, Bacterial

KW - Formaldehyde

KW - Humans

KW - Paraffin Embedding

KW - Pneumocystis jirovecii

KW - Real-Time Polymerase Chain Reaction

KW - Sensitivity and Specificity

KW - Tissue Fixation

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U2 - 10.1016/j.yexmp.2015.03.021

DO - 10.1016/j.yexmp.2015.03.021

M3 - Article

C2 - 25779023

SN - 0014-4800

VL - 98

SP - 390

EP - 392

JO - Experimental and molecular pathology

JF - Experimental and molecular pathology

IS - 3

ER -

Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples

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Keywords

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