Incorporating microarray assessment of HER2 status in clinical practice supports individualised therapy in early-stage breast cancer.

Accurate determination of human epidermal growth factor receptor-2 (HER2) status is essential for optimal selection of breast cancer patients for gene targeted therapy. The analytical performance of microarray analysis using TargetPrint for assessment of HER2 status was evaluated in 138 breast tumours, including 41 fresh and 97 formalin-fixed paraffin embedded (FFPE) specimens. Reflex testing using immunohistochemistry/in situ hybridization (IHC/ISH) in four discordant cases confirmed the TargetPrint results, achieving 100% agreement regardless of whether fresh tissue or FFPE specimens were used. One equivocal IHC/ISH case was classified as HER2-positive based on the microarray result. The proven clinical utility in resolving equivocal and borderline cases justifies modification of the testing algorithm under these circumstances, to obtain a definitive positive or negative test result with the use of microarrays. Determination of HER2 status across three assay platforms facilitated improved quality assurance and led to a higher level of confidence on which to base treatment decisions.

Comparison of the quality of smears in transbronchial fine-needle aspirates using two staining methods for rapid on-site evaluation.

Transbronchial needle aspiration (TBNA) via flexible bronchoscopy is a well-established sampling modality for lung masses. The procedure is useful in the diagnosis of neoplastic and non-neoplastic lesions as well as for staging of bronchogenic carcinoma. Rapid on-site evaluation (ROSE) adds value as it has the advantage of triaging material during the procedure so avoiding a battery of investigations. Frequently used rapid stains are the modified Wright-Giemsa water-based stain (WG-ROSE) and the alcohol-based modified Papanicolaou stain (Pap-ROSE). Final review of laboratory-based Giemsa and Pap stains supplemented by ancillary investigations is essential for quality assurance. To investigate whether and how ROSE influenced the quantity and quality of the material submitted to the laboratory we randomized 126 patients to WG-ROSE, requiring only one pathologist on-site, or combined WG- and Pap-ROSE, requiring an additional person on-site to assist with staining. In those patients with positive TBNA we graded the laboratory-based slides of the first pass containing diagnostic material into insufficient, suspicious, adequate and excellent. The first diagnostic pass was found after 3.06 ± 1.94 (SD) passes and 3.13 ± 2.16 passes with WG-ROSE and combined ROSE (P = 0.87), respectively. Following WG-ROSE and combined ROSE 69% and 71.1% (P = 0.509) of slides were diagnostic (adequate or excellent) on laboratory-based Giemsa stains, and 93.3% and 100% (P = 0.134) were scored adequate or excellent on laboratory-based Pap stains. We concluded that the less costly and labour intensive WG-ROSE procedure is adequate for TBNA. This has cost implications especially in resource poor settings.

APRI: a simple bedside marker for advanced fibrosis that can avoid liver biopsy in patients with NAFLD/NASH.

BACKGROUND: Non-alcoholic steatohepatitis (NASH) can lead to cirrhosis and hepatocellular carcinoma. The NASH fibrosis score (NFS) has proven to be a reliable, non-invasive marker for prediction of advanced fibrosis. Aspartate aminotransferase-to-platelet ratio index (APRI) is a simpler calculation than NFS, but has never been studied in patients with non-alcoholic fatty liver disease (NAFLD). AIM: To validate APRI as a non-invasive marker of liver fibrosis in subjects with NAFLD to be used in clinical practice. DESIGN/METHODS: The cohort consisted of 111 patients with histological diagnoses of NAFLD. The biopsy samples were staged and graded according to the NASH clinical research network (CRN) criteria. These were grouped into fatty liver disease (FLD), NASH, no/mild fibrosis, and advanced fibrosis. The sensitivity and specificity of APRI were compared with NFS and aspartate aminotransferase-to-alanine aminotransferase (AST/ALT) ratio. RESULTS: The APRI was significantly higher in the advanced fibrosis group. The area under receiver operating characteristic (ROC) curve for APRI was 0.85 with an optimal cut-off of 0.98, giving a sensitivity of 75% and a specificity of 86%. The NFS was significantly lower in the advanced fibrosis group. The ROC for NFS gave an area under curve (AUC) of 0.77 and a cut-off value of -1.3 with a sensitivity of 76% and specificity of 69%. The positive predictive value for APRI was 54% as opposed to 34% for NFS. The negative predictive value was 93% for APRI and 94% for NFS. CONCLUSION: APRI compared favourably to NFS and was superior to AST/ALT for the prediction of advanced fibrosis. We therefore propose the use of APRI in a new algorithm for the detection of advanced fibrosis.

Fine needle aspiration biopsy and flow cytometry in the diagnosis of lymphoma.

UNLABELLED: Fine needle aspiration biopsy (FNAB) is emerging as a technique of potential value in the diagnosis of benign and malignant lesions in areas such as the breast and thyroid gland. Its place in distinguishing reactive from neoplastic lymphoid proliferations, when compared to the established practice of excision biopsy and histopathology, continues to undergo evaluation. Morphology alone discriminates poorly between atypical or lymphoproliferative disorders as seen in the presence of Epstein-Barr or human immunodeficiency virus. Furthermore the polymorphic populations of follicular lymphoma may mimic reactive changes. In addition previous classifications of these tumours using working formulation or Kiel classification relied heavily on architecture, which is a feature not reflected in cytology smears. The World Health Organisation approach includes clinical features, immunophenotyping and cytogenetic profiles to define neoplasms of immunohaematopoietic tissues. Flow cytometry on fine needle aspiration biopsy offers additional advantages in being rapid and objective in quantitatively as well as qualitatively documenting cell surface characteristics. All patients referred for this procedure to Tygerberg Academic Hospital with suspected nodal or extranodal lymphoma between January 2002 and December 2004 were analysed. In each case flow cytometry and cytomorphology were correlated with histopathology on tissue biopsy, bone marrow examination and clinical follow-up for confirmation of diagnosis. Results of the 124 cases were tabulated and statistically processed. Eighty-one met the inclusion criteria, thirteen (16.1%) were not malignant, two (2.5%) were falsely negative, two (2.5%) were equivocal needing histology and in the remaining sixty-four (79%) diagnosis was achieved. SUMMARY: Fine needle aspiration coupled with flow cytometry can reliably distinguish between nodal and extranodal neoplastic B-cell population. It is concluded that appropriate use, in a collaborative multidisciplinary setting, may eliminate the need for surgical procedures in many cases. CONCLUSION: These advances are not widely recognised and this is particularly true in South Africa. Accordingly, such an approach has been prospectively evaluated in the Western Cape showing that the combination of ready availability and diagnostic accuracy, after an initial learning curve, allow accurate characterisation of haematologic malignancies so that excision biopsy may be reserved for specific further studies to provide data not available from this less invasive procedure.

Ultrasound-assisted transthoracic biopsy: cells or sections?

Physicians increasingly use transthoracic ultrasound (US) as an aid for diagnostic procedures. At the bedside, US helps to visualize neoplasms in the chest wall, pleura, peripheral lung, and anterior mediastinum involving or abutting the pleura. Histology specimens from cutting-needle biopsies have been shown to be safe and effective. This prospective study determined the yield and safety of US-guided fine-needle aspiration biopsy (FNAB) as a first-line bedside investigation. We recruited 97 consecutive patients, and of these, 85 underwent both cutting-needle biopsy and FNAB. These were adequate for diagnosis in 81.2% and 80% of cases, respectively, with a combined yield of 90%. Measured with a novel semiquantitative score, FNAB allowed a diagnosis with fewer special investigations than cutting biopsy. US-guided FNAB by pulmonologists performed best in lung carcinoma and can be recommended as a first-line investigation in patients with a high clinical suspicion of this diagnosis.

Transbronchial needle aspirates: comparison of two preparation methods.

STUDY OBJECTIVES: Transbronchial needle aspiration has evolved as a key bronchoscopic sampling method. Specimen handling and preparation are underrated yet crucial aspects of the technique. This study was designed to identify which of two widely practiced sample preparation methods has a higher yield. DESIGN: Prospective comparison of two diagnostic methods. SETTING: Tertiary academic hospital. PATIENTS: Consecutive patients undergoing transbronchial needle aspiration. INTERVENTIONS: Transbronchial aspirates were obtained pairwise. One specimen was placed directly onto a slide and smears were prepared on site (ie, the direct technique), and the other specimen was deposited into a vial containing 95% alcohol and further prepared in the laboratory (ie, the fluid technique). In total, 282 pairs of samples were aspirated from 145 target sites (paratracheal, 10 sites; tracheobronchial, 101 sites; hilar, 17 sites; endobronchial or peripheral, 17 sites). MEASUREMENTS AND RESULTS: The measured outcome was the presence of diagnostic material at the final laboratory assessment. At least one diagnostic aspirate was obtained in 66% of 86 investigated patients (small cell lung cancer, 18 patients; non-small cell lung cancer, 47 patients; other diagnoses, 21 patients). The direct technique had a better yield overall than the fluid technique (positive aspirates, 36.2% vs 12.4%, respectively; p < 0.01), as well as after stratification for tumor type and for anatomic site. CONCLUSION: The direct technique is superior to the fluid technique for the preparation of transbronchial needle aspirates.

Utility of rapid on-site evaluation of transbronchial needle aspirates.

BACKGROUND: Rapid on-site evaluation has been proposed as a method to improve the yield of transbronchial needle aspiration. OBJECTIVES: This study investigated whether on-site analysis facilitates routine diagnostic bronchoscopy in terms of sampling, yield and cost. METHODS: Patients with lesions accessible for transbronchial needle aspiration on computed tomography were investigated. A cytopathologist screened the needle aspirates on site for the presence of diagnostic material. The bronchoscopic sampling process was adjusted according to the results. In 90 consecutive patients with neoplastic disease (n=70; 78%), non-neoplastic disease (n=16; 18%) or undiagnosed lesions (n=4; 4%) we aspirated 162 lung tumours or lymph node sites (mediastinal: 7%; tracheobronchial: 68%; other: 25%). In 90 consecutive patients with neoplastic disease (n=70; 78%), non-neoplastic disease (n=16; 18%) or undiagnosed lesions (n=4; 4%) we aspirated 162 lung lesions (paratracheal tumours or lymph nodes: 7%; tracheobronchial lymph nodes: 68%; other: 25%). RESULTS: The diagnostic yield of needle aspiration was 77 and 25% in patients with neoplastic and non-neoplastic lesions, respectively. Sampling could be terminated in 64% of patients after needle aspiration had been performed as the only diagnostic modality, and on-site analysis identified diagnostic material from the first site aspirated in 50% of patients. Only in 2 patients (2%) diagnostic aspirates were not recognized on site. On-site analysis was cost effective due to savings for disposable diagnostic tools, which exceeded the extra expense for the on-site cytology service provided. CONCLUSIONS: Rapid on-site analysis of transbronchial aspirates is a highly useful, accurate and cost-effective addition to routine diagnostic bronchoscopy.