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Reliable EGFR mutation testing in ultrasound-guided supraclavicular lymph node fine-needle aspirates: a cohort study with diagnostic performance analysis.

Awwad A, Tiwari S, Sovani V, Baldwin DR, Kumaran M - BMJ Open Respir Res (2015)

Bottom Line: Ultrasound (US)-guided fine-needle aspiration (FNA) cytology is regarded as an effective diagnostic tool in small size lymph nodes (LNs) and impalpable positron emission tomography detected nodes.The remaining 57 patients had further investigations; 45 reconfirmed the US-guided FNA diagnosis.No complications were recorded.

View Article: PubMed Central - PubMed

Affiliation: Radiology Department , Nottingham University Hospitals NHS Trust, Queen's Medical Centre , Nottingham, Nottinghamshire , UK.

ABSTRACT

Introduction: 15-30% of patients with lung cancer will have supraclavicular and cervical lymphadenopathy (SCLN). Ultrasound (US)-guided fine-needle aspiration (FNA) cytology is regarded as an effective diagnostic tool in small size lymph nodes (LNs) and impalpable positron emission tomography detected nodes. We evaluated our diagnostic service performance in relation to the adequacy of samples for epidermal growth factor receptor (EGFR) mutation.

Methods: Retrospective data analysis from electronic records, searching for all suspected lung cancer referrals that underwent US of the neck±FNA, over a continuous period of 4 years.

Results: Of 306 cases with suspected lung cancer referred to our department for US FNA of SCLN, 228 patients underwent the procedure. Of the remaining 78 patients, LNs were not detected in 52 cases and appeared benign in 26. Cytological diagnosis was established in 171 patients (75%) for treatment decisions without further investigations. The remaining 57 patients had further investigations; 45 reconfirmed the US-guided FNA diagnosis. The average LN size was 12.9 mm, and positive cytology was obtained in LNs ranging from 3 to 45 mm. Of 57 adenocarcinoma cases, 34 were tested for EGFR confirming 4 positive, 25 negative and 5 insufficient samples. No complications were recorded.

Conclusions: US-guided FNA of SCLN remains an important diagnostic tool in lung cancer. Adequate tissue can be obtained for reliable diagnosis from LNs and for EGFR mutational analysis, without the need for more invasive and expensive investigations in more than 80% of cases.

No MeSH data available.


Related in: MedlinePlus

(A–C) Electron microscopic images obtained from two different cases, (A) a cluster of large malignant cells with cytoplasmic mucin vacuoles consistent with metastatic adenocarcinoma (Pap stain, ×40 magnifications). (B) Large malignant cells with vesicular nuclei and prominent nucleoli consistent with metastatic non-small cell carcinoma (Pap stain, ×40 magnifications). (C) Thyroid transcription factor 1 (TTF1, ×20 magnification) immunostaining slide showing nuclear positivity confirming metastatic adenocarcinoma.
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BMJRESP2014000075F2: (A–C) Electron microscopic images obtained from two different cases, (A) a cluster of large malignant cells with cytoplasmic mucin vacuoles consistent with metastatic adenocarcinoma (Pap stain, ×40 magnifications). (B) Large malignant cells with vesicular nuclei and prominent nucleoli consistent with metastatic non-small cell carcinoma (Pap stain, ×40 magnifications). (C) Thyroid transcription factor 1 (TTF1, ×20 magnification) immunostaining slide showing nuclear positivity confirming metastatic adenocarcinoma.

Mentions: In current practice, the most reported techniques to test for EGFR mutations are reliant on PCR.712 Three highly sensitive methods are referred to in literature which would either employ the Peptide Nucleic Acid—Locked Nucleic Acid PCR clamp, the Cycleave method and/or the PCR invader.913 All are capable of yielding DNA positional defects with a 1% ratio of cancer cells in a specimen.14 However, quantification and qualitative analysis of extracted DNA samples in our cohort is beyond the remit of our report. In our institution laboratories, we use pyrosequencing, which is an assay based on nested PCR for the characterisation of these mutations on formalin-fixed and paraffin-embedded tumour tissue (added advantage) as well as cytospins from FNAs. The yield of malignant cells on aspirated material is often variable. All samples were first analysed by specialist pulmonary pathologists to classify tumours according to the WHO classification. Where the diagnosis of adenocarcinoma was not possible on the initial stains, samples were spun down to make into a cell block. The latter was found more suitable for immunohistochemical analysis and EGFR testing, especially as it is more predictive of the percentage of malignant cells in paucicellular samples. DNA was extracted using the manufacturer's guidelines and subjected to nested PCR to achieve the required amplification (see figure 2A–C for more details).


Reliable EGFR mutation testing in ultrasound-guided supraclavicular lymph node fine-needle aspirates: a cohort study with diagnostic performance analysis.

Awwad A, Tiwari S, Sovani V, Baldwin DR, Kumaran M - BMJ Open Respir Res (2015)

(A–C) Electron microscopic images obtained from two different cases, (A) a cluster of large malignant cells with cytoplasmic mucin vacuoles consistent with metastatic adenocarcinoma (Pap stain, ×40 magnifications). (B) Large malignant cells with vesicular nuclei and prominent nucleoli consistent with metastatic non-small cell carcinoma (Pap stain, ×40 magnifications). (C) Thyroid transcription factor 1 (TTF1, ×20 magnification) immunostaining slide showing nuclear positivity confirming metastatic adenocarcinoma.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488608&req=5

BMJRESP2014000075F2: (A–C) Electron microscopic images obtained from two different cases, (A) a cluster of large malignant cells with cytoplasmic mucin vacuoles consistent with metastatic adenocarcinoma (Pap stain, ×40 magnifications). (B) Large malignant cells with vesicular nuclei and prominent nucleoli consistent with metastatic non-small cell carcinoma (Pap stain, ×40 magnifications). (C) Thyroid transcription factor 1 (TTF1, ×20 magnification) immunostaining slide showing nuclear positivity confirming metastatic adenocarcinoma.
Mentions: In current practice, the most reported techniques to test for EGFR mutations are reliant on PCR.712 Three highly sensitive methods are referred to in literature which would either employ the Peptide Nucleic Acid—Locked Nucleic Acid PCR clamp, the Cycleave method and/or the PCR invader.913 All are capable of yielding DNA positional defects with a 1% ratio of cancer cells in a specimen.14 However, quantification and qualitative analysis of extracted DNA samples in our cohort is beyond the remit of our report. In our institution laboratories, we use pyrosequencing, which is an assay based on nested PCR for the characterisation of these mutations on formalin-fixed and paraffin-embedded tumour tissue (added advantage) as well as cytospins from FNAs. The yield of malignant cells on aspirated material is often variable. All samples were first analysed by specialist pulmonary pathologists to classify tumours according to the WHO classification. Where the diagnosis of adenocarcinoma was not possible on the initial stains, samples were spun down to make into a cell block. The latter was found more suitable for immunohistochemical analysis and EGFR testing, especially as it is more predictive of the percentage of malignant cells in paucicellular samples. DNA was extracted using the manufacturer's guidelines and subjected to nested PCR to achieve the required amplification (see figure 2A–C for more details).

Bottom Line: Ultrasound (US)-guided fine-needle aspiration (FNA) cytology is regarded as an effective diagnostic tool in small size lymph nodes (LNs) and impalpable positron emission tomography detected nodes.The remaining 57 patients had further investigations; 45 reconfirmed the US-guided FNA diagnosis.No complications were recorded.

View Article: PubMed Central - PubMed

Affiliation: Radiology Department , Nottingham University Hospitals NHS Trust, Queen's Medical Centre , Nottingham, Nottinghamshire , UK.

ABSTRACT

Introduction: 15-30% of patients with lung cancer will have supraclavicular and cervical lymphadenopathy (SCLN). Ultrasound (US)-guided fine-needle aspiration (FNA) cytology is regarded as an effective diagnostic tool in small size lymph nodes (LNs) and impalpable positron emission tomography detected nodes. We evaluated our diagnostic service performance in relation to the adequacy of samples for epidermal growth factor receptor (EGFR) mutation.

Methods: Retrospective data analysis from electronic records, searching for all suspected lung cancer referrals that underwent US of the neck±FNA, over a continuous period of 4 years.

Results: Of 306 cases with suspected lung cancer referred to our department for US FNA of SCLN, 228 patients underwent the procedure. Of the remaining 78 patients, LNs were not detected in 52 cases and appeared benign in 26. Cytological diagnosis was established in 171 patients (75%) for treatment decisions without further investigations. The remaining 57 patients had further investigations; 45 reconfirmed the US-guided FNA diagnosis. The average LN size was 12.9 mm, and positive cytology was obtained in LNs ranging from 3 to 45 mm. Of 57 adenocarcinoma cases, 34 were tested for EGFR confirming 4 positive, 25 negative and 5 insufficient samples. No complications were recorded.

Conclusions: US-guided FNA of SCLN remains an important diagnostic tool in lung cancer. Adequate tissue can be obtained for reliable diagnosis from LNs and for EGFR mutational analysis, without the need for more invasive and expensive investigations in more than 80% of cases.

No MeSH data available.


Related in: MedlinePlus