EGFR mutational genotyping of liquid based cytology samples obtained via fine needle aspiration (FNA) at endobronchial ultrasound of non-small cell lung cancer (NSCLC)

Jordan P. Reynolds, Raymond R. Tubbs, Eugen C. Minca, Stephen MacNamara, Francisco A. Almeida, Patrick C. Ma, Nathan A. Pennell, Joseph C. Cicenia

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Objectives: Epidermal growth factor receptor (. EGFR) gene mutation status should be determined in all patients with advanced, non-squamous non-small cell lung carcinoma (NSCLC) to guide targeted therapy with EGFR tyrosine kinase inhibitors. EGFR mutations are commonly tested by Sanger sequencing or allele specific polymerase chain reaction (ASPCR) on formalin-fixed paraffin-embedded (FFPE) samples including cell blocks (CB) that may fail due to absence of tumor cells. The cell pellet from cytology specimens obtained at the time of endobronchial guided ultrasound fine needle aspiration (EBUS FNA) (EBUS-TBNA, transbronchial needle aspiration) represents an alternative resource for additional tissue. Here we demonstrate the utility of using the FNA cell pellet versus for the detection of EGFR mutations in NSCLC. Materials and methods: For internal validation, 39 cytology samples from patients with NSCLC referred for EGFR testing were analyzed using the EGFR rotor-gene Q (RGQ) PCR assay (Qiagen).Thereafter, a consecutive series of 228 EBUS FNA samples were tested. Results: The ASPCR assay demonstrated acceptable intra-assay, inter-assay and inter-lot reproducibility, sensitivity, and specificity. For the consecutive series, only 6/228 (2.6%) failed analysis (5 due to insufficient DNA yield). Of 228 EBUS FNA cell pellets tested 32 (14.0%) demonstrated clinically relevant mutations. Results and conclusion: ASPCR can reliably detect EGFR gene mutations in FNA preparations from patients with NSCLC obtained at EBUS.

Original languageEnglish (US)
Pages (from-to)158-163
Number of pages6
JournalLung Cancer
Volume86
Issue number2
DOIs
StatePublished - Nov 1 2014

Fingerprint

Fine Needle Biopsy
Non-Small Cell Lung Carcinoma
Cell Biology
erbB-1 Genes
Mutation
Polymerase Chain Reaction
Alleles
Paraffin
Protein-Tyrosine Kinases
Formaldehyde
Needles
Sensitivity and Specificity
DNA
Neoplasms
Therapeutics

All Science Journal Classification (ASJC) codes

  • Oncology
  • Pulmonary and Respiratory Medicine
  • Cancer Research

Cite this

Reynolds, Jordan P. ; Tubbs, Raymond R. ; Minca, Eugen C. ; MacNamara, Stephen ; Almeida, Francisco A. ; Ma, Patrick C. ; Pennell, Nathan A. ; Cicenia, Joseph C. / EGFR mutational genotyping of liquid based cytology samples obtained via fine needle aspiration (FNA) at endobronchial ultrasound of non-small cell lung cancer (NSCLC). In: Lung Cancer. 2014 ; Vol. 86, No. 2. pp. 158-163.
@article{eeb98dca23b14f4c8208fc4dd69062ab,
title = "EGFR mutational genotyping of liquid based cytology samples obtained via fine needle aspiration (FNA) at endobronchial ultrasound of non-small cell lung cancer (NSCLC)",
abstract = "Objectives: Epidermal growth factor receptor (. EGFR) gene mutation status should be determined in all patients with advanced, non-squamous non-small cell lung carcinoma (NSCLC) to guide targeted therapy with EGFR tyrosine kinase inhibitors. EGFR mutations are commonly tested by Sanger sequencing or allele specific polymerase chain reaction (ASPCR) on formalin-fixed paraffin-embedded (FFPE) samples including cell blocks (CB) that may fail due to absence of tumor cells. The cell pellet from cytology specimens obtained at the time of endobronchial guided ultrasound fine needle aspiration (EBUS FNA) (EBUS-TBNA, transbronchial needle aspiration) represents an alternative resource for additional tissue. Here we demonstrate the utility of using the FNA cell pellet versus for the detection of EGFR mutations in NSCLC. Materials and methods: For internal validation, 39 cytology samples from patients with NSCLC referred for EGFR testing were analyzed using the EGFR rotor-gene Q (RGQ) PCR assay (Qiagen).Thereafter, a consecutive series of 228 EBUS FNA samples were tested. Results: The ASPCR assay demonstrated acceptable intra-assay, inter-assay and inter-lot reproducibility, sensitivity, and specificity. For the consecutive series, only 6/228 (2.6{\%}) failed analysis (5 due to insufficient DNA yield). Of 228 EBUS FNA cell pellets tested 32 (14.0{\%}) demonstrated clinically relevant mutations. Results and conclusion: ASPCR can reliably detect EGFR gene mutations in FNA preparations from patients with NSCLC obtained at EBUS.",
author = "Reynolds, {Jordan P.} and Tubbs, {Raymond R.} and Minca, {Eugen C.} and Stephen MacNamara and Almeida, {Francisco A.} and Ma, {Patrick C.} and Pennell, {Nathan A.} and Cicenia, {Joseph C.}",
year = "2014",
month = "11",
day = "1",
doi = "10.1016/j.lungcan.2014.09.003",
language = "English (US)",
volume = "86",
pages = "158--163",
journal = "Lung Cancer",
issn = "0169-5002",
publisher = "Elsevier Ireland Ltd",
number = "2",

}

EGFR mutational genotyping of liquid based cytology samples obtained via fine needle aspiration (FNA) at endobronchial ultrasound of non-small cell lung cancer (NSCLC). / Reynolds, Jordan P.; Tubbs, Raymond R.; Minca, Eugen C.; MacNamara, Stephen; Almeida, Francisco A.; Ma, Patrick C.; Pennell, Nathan A.; Cicenia, Joseph C.

In: Lung Cancer, Vol. 86, No. 2, 01.11.2014, p. 158-163.

Research output: Contribution to journalArticle

TY - JOUR

T1 - EGFR mutational genotyping of liquid based cytology samples obtained via fine needle aspiration (FNA) at endobronchial ultrasound of non-small cell lung cancer (NSCLC)

AU - Reynolds, Jordan P.

AU - Tubbs, Raymond R.

AU - Minca, Eugen C.

AU - MacNamara, Stephen

AU - Almeida, Francisco A.

AU - Ma, Patrick C.

AU - Pennell, Nathan A.

AU - Cicenia, Joseph C.

PY - 2014/11/1

Y1 - 2014/11/1

N2 - Objectives: Epidermal growth factor receptor (. EGFR) gene mutation status should be determined in all patients with advanced, non-squamous non-small cell lung carcinoma (NSCLC) to guide targeted therapy with EGFR tyrosine kinase inhibitors. EGFR mutations are commonly tested by Sanger sequencing or allele specific polymerase chain reaction (ASPCR) on formalin-fixed paraffin-embedded (FFPE) samples including cell blocks (CB) that may fail due to absence of tumor cells. The cell pellet from cytology specimens obtained at the time of endobronchial guided ultrasound fine needle aspiration (EBUS FNA) (EBUS-TBNA, transbronchial needle aspiration) represents an alternative resource for additional tissue. Here we demonstrate the utility of using the FNA cell pellet versus for the detection of EGFR mutations in NSCLC. Materials and methods: For internal validation, 39 cytology samples from patients with NSCLC referred for EGFR testing were analyzed using the EGFR rotor-gene Q (RGQ) PCR assay (Qiagen).Thereafter, a consecutive series of 228 EBUS FNA samples were tested. Results: The ASPCR assay demonstrated acceptable intra-assay, inter-assay and inter-lot reproducibility, sensitivity, and specificity. For the consecutive series, only 6/228 (2.6%) failed analysis (5 due to insufficient DNA yield). Of 228 EBUS FNA cell pellets tested 32 (14.0%) demonstrated clinically relevant mutations. Results and conclusion: ASPCR can reliably detect EGFR gene mutations in FNA preparations from patients with NSCLC obtained at EBUS.

AB - Objectives: Epidermal growth factor receptor (. EGFR) gene mutation status should be determined in all patients with advanced, non-squamous non-small cell lung carcinoma (NSCLC) to guide targeted therapy with EGFR tyrosine kinase inhibitors. EGFR mutations are commonly tested by Sanger sequencing or allele specific polymerase chain reaction (ASPCR) on formalin-fixed paraffin-embedded (FFPE) samples including cell blocks (CB) that may fail due to absence of tumor cells. The cell pellet from cytology specimens obtained at the time of endobronchial guided ultrasound fine needle aspiration (EBUS FNA) (EBUS-TBNA, transbronchial needle aspiration) represents an alternative resource for additional tissue. Here we demonstrate the utility of using the FNA cell pellet versus for the detection of EGFR mutations in NSCLC. Materials and methods: For internal validation, 39 cytology samples from patients with NSCLC referred for EGFR testing were analyzed using the EGFR rotor-gene Q (RGQ) PCR assay (Qiagen).Thereafter, a consecutive series of 228 EBUS FNA samples were tested. Results: The ASPCR assay demonstrated acceptable intra-assay, inter-assay and inter-lot reproducibility, sensitivity, and specificity. For the consecutive series, only 6/228 (2.6%) failed analysis (5 due to insufficient DNA yield). Of 228 EBUS FNA cell pellets tested 32 (14.0%) demonstrated clinically relevant mutations. Results and conclusion: ASPCR can reliably detect EGFR gene mutations in FNA preparations from patients with NSCLC obtained at EBUS.

UR - http://www.scopus.com/inward/record.url?scp=84908404380&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84908404380&partnerID=8YFLogxK

U2 - 10.1016/j.lungcan.2014.09.003

DO - 10.1016/j.lungcan.2014.09.003

M3 - Article

C2 - 25263855

AN - SCOPUS:84908404380

VL - 86

SP - 158

EP - 163

JO - Lung Cancer

JF - Lung Cancer

SN - 0169-5002

IS - 2

ER -