Integral Part Of Diagnostic Workup Research Articles

Formalin Sediment Cytology of Small Biopsy Samples: An Adjunct to Histopathology

Introduction: Cytological evaluation of malignant neoplasms forms an integral part of diagnostic work-up in any malignant disorders. Sediment cytology or more aptly called as biopsy sediment cytology is a technique that involves study of smears prepared from sediment of biopsy specimen fixatives. The preliminary diagnosis on sediment cytology helps in planning and early initiation of treatment. Aim: To evaluate the cytomorphological features of sediment cytology of small biopsy samples and compare the same with histopathological diagnosis. Materials and Methods: A cross-sectional study was conducted over a period of two months from July 2021 to August 2021 at ESIC Medical College and PGIMSR, Bengaluru, Karnataka, India. A total of 51 samples were included for the study. The sediments of biopsy specimens were collected in a test tube, centrifuged at 2500 rpm for 10 minutes and smears were prepared. Stained smears were evaluated for presence and morphology of cells. The morphology was compared with histopathological diagnosis of biopsy specimens. The concordance rate was assessed by Pearson’s correlation coefficient in non neoplastic/benign lesions and malignant lesions separately. A p value of <0.05 was taken to be statistically significant. Results: Biopsies from gastrointestinal system accounted for the maximum number of cases (16 cases, 31.4%), followed by biopsies from oral cavity (15 cases, 29.4%) and female genital tract (11 cases, 21.5%). Sediment cytology yielded diagnostic material in 46 cases (90.2%). The diagnostic yield was 90.2% with an overall concordance of 60.8%. The sensitivity, specificity and diagnostic accuracy was 65.12%, 100% and 60.8% respectively. The concordance rate in non neoplastic/benign lesions was r=0.99, with p=0.0001, whereas with respect to malignant lesions, concordance rate was r=0.86 with p=0.0003. Conclusion: Cytological evaluation plays an important role in early and effective planning of appropriate treatment. Biopsy sediment cytology although an adjunct to histopathology has practical utility and fulfills the desired role of any cytological sample in providing a provisional diagnosis.

Endleukemia Assay v1: Enabling NGS-Based Comprehensive Routine Molecular Profiling of Leukemias in Routine Clinical Care

Background: Molecular biomarkers provide diagnostic, prognostic, predictive and minimal residual disease (MRD) information and constitute an integral part of diagnostic workup of hematologic neoplasms. Since a variety of different hematologic disorders can often show non-specific clinical symptoms as well as laboratory findings, the initial diagnostic workup needs to be optimized for timely and efficient coverage of a broad range of differential diagnoses. We developed a customized 81-gene panel, EndLeukemia Assay v1, for comprehensive genetic characterization of disorders that include bone marrow (BM) and peripheral blood (PB). The biomarkers on the panel cover genetic alterations in myeloid neoplasms (acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN), myelodysplastic syndrome/myeloproliferative neoplasms (MDS/MPN), aplastic anemia (AA), acute lymphoblastic leukemia (ALL), hairy cell leukemia, T-large granular lymphocytic leukemia (T-LGL) and T-prolymphocytic leukemia (T-PLL).Materials and Methods: Genomic DNA was isolated from fresh BM aspirate/PB or formalin-fixed paraffin-embedded (FFPE) BM clot specimens. Either entire coding sequences or limited exons/hotspots for 81-gene target genes were captured from 250 ng DNA using the hybridization-based HaloPlexTM target enrichment system (Agilent Technologies, Santa Clara, CA). These genes include A NKRD26, ASXL1, ASXL2, BCOR, BCORL1, BRAF, BRINP3, CALR, CBL, CBLB, CBLC, CEBPA, CREBBP, CRLF2, CSF3R, CUX1, DDX41, DNMT3A, EED, ELANE, ETNK1, ETV6, EZH2, FBXW7, FLT3, GATA1, GATA2, GFI1, GNAS, HNRNPK, HRAS, IDH1, IDH2, IKZF1, IL2RG, IL7R, JAK1, JAK2, JAK3, KDM6A, KIT, KMT2A, KRAS, MAP2K1, MPL, NF1, NOTCH1, NPM1, NRAS, PAX5, PHF6, PIGA, PML, PRPF40B, PTEN, PTPN11, RAD21, RARA, RUNX1, SETBP1, SF1, SF3A1, SF3B1, SH2B3, SMC1A, SMC3, SRSF2, STAG1, STAG2, STAT3, STAT5A, STAT5B, SUZ12, TERC, TERT, TET2, TP53, U2AF1, U2AF2, WT1, and ZRSR2. The panel design incorporated molecular barcodes to improve accuracy and sensitivity. Bidirectional paired-end sequencing was performed using the Illumina Miseq platform (Illumina, San Diego, CA). Variant calling was performed using Agilent SureCall software v3.5 and an in-house developed bioinformatics pipeline. Incorporation of single nucleotide polymorphisms (SNP) allows detection of sample identification errors and improves patient safety.Results: During the validation studies, high concordance was observed for missense mutations, insertions/deletions (indels), and selected copy number aberrations (CNAs) when compared with orthogonal methods. Under standardized conditions, a minimum average coverage depth of 3000x is achieved across the entire target region. The analytical sensitivity of the assay was established at 5% using a per base coverage cut-off of 250x. Of note, >90% of the CEBPA coding sequence, which is challenging to sequence due to a high GC component, was covered at >250x coverage depth. Copy number alterations including KMT2A partial tandem duplications (KMT2A -PTD) and IKZF1 deletions were detected successfully. Initial validation of FFPE tissue shows 100% concordance for all mutation calls for matched fresh and FFPE sample pairs. The ability to provide critical diagnostic and prognostic information across mutation hotspots from FFPE tissue if a fresh PB/BM sample is not available improves patient care. The top 25 mutations in the first 500 patient samples across different clinical indications are shown in the figure below. In addition, efficient panel design allowed uniform testing of follow-up bone marrow samples for the detection of MRD and clonal evolution.Conclusion: EndLeukemia assay v1 allowed comprehensive molecular profiling of a variety of hematologic neoplasms in routine clinical testing. Routine molecular biomarker profiling can be facilitated by incorporating smart design features in clinical NGS panels. [Display omitted] DisclosuresKantarjian:Delta-Fly Pharma: Research Funding; Novartis: Research Funding; ARIAD: Research Funding; Pfizer: Research Funding; Amgen: Research Funding; Bristol-Meyers Squibb: Research Funding.

Does 68Ga-DOTA-NOC-PET/CT impact staging and therapeutic decision making in pulmonary carcinoid tumors?

Purpose of this study was to assess the utility of Ga-DOTA-NOC-PET/computed tomography (CT) (Ga-NOC-PET) in tumor detection, pathological differentiation and baseline staging of pulmonary carcinoids as well as to study its impact on therapeutic decision making. Patients who underwent a Ga-NOC-PET for initial evaluation of bronchopulmonary carcinoid tumors from August 2014 to December 2019 were included. Detection rate of Ga-NOC-PET for the primary lesion was calculated by visual estimation of tracer uptake as per Krenning score. SUVmax of typical and atypical carcinoid tumors was measured and difference compared using nonparametric statistical tests. Proportion of patients with distant metastases was also calculated and its impact on intended treatment was assessed. Imaging, histopathology and treatment details of 119 patients were available for analysis. Majority of tumors had an endobronchial location (74.7%) and showed histopathologic features of typical carcinoid (82.3%). Ga-NOC-PET showed a detection rate/sensitivity of 92.4%. Oncocytic variant on histopathology and smaller tumor size accounted for majority of negative results. Typical carcinoids showed significantly higher SUVmax than atypical tumors (median SUVmax 38.4 vs. 15.7, P = 0.002). Metastases to distant sites outside the thorax were seen in 14 patients (11.7%), primarily in liver and bones changing the intent of treatment from surgery to systemic therapy. Ga-NOC-PET detects asymptomatic distant metastatic disease in a sizeable number of patients (11.7%) with pulmonary carcinoid and thus contribute to clinical management by precluding futile surgeries. It shows a high sensitivity for tumor detection and can help differentiate between typical and atypical carcinoid variants by virtue of their variable tracer uptake. PET/CT using Ga-labeled DOTA peptides should be an integral part of diagnostic workup of patients with lung carcinoid.

Multimodality Imaging Findings of a Renal Aspergilloma.

Renal aspergillosis is a rare infection that usually occurs in persons with a predisposition for this condition. Its differential diagnosis includes primary and metastatic renal malignancies, pyelonephritis and secondary abscess formation, granulomatous disorders, and renal infarction. We aim to stress the role of multimodality imaging and percutaneous biopsy in the diagnosis of this condition. We present diffusion weighted imaging (DWI) and positron emission tomography-computed tomography (PET-CT) findings in addition to conventional imaging modalities in a 55-year-old man with secondary renal aspergilloma. Radiological imaging methods are an integral part of diagnostic workup for renal aspergillosis. A definitive diagnosis is made by histopathological and/or microbiological examination of the material obtained via percutaneous biopsy under guidance of imaging methods.