Frequency and Aberrant CD Marker Profile of Acute Leukaemias.

OBJECTIVE: To determine the frequency of different types of acute leukaemia and their subtypes along with associated aberrant CD markers. STUDY DESIGN: Descriptive study. Place and Duration of the Study: Department of Immunology Armed Forces Institute of Pathology, National University of Medical Sciences, Rawalpindi, Pakistan, from November 2021 to October 2023. METHODOLOGY: All samples received for flow cytometric immunophenotyping with suspicion of acute leukaemia were included in the study. Cells were stained with fluorochrome labelled monoclonal antibodies against lineage-specific cluster of differentiation (CD) markers through a lyse-wash procedure. Acquisition and analysis were done using multi-parameter BD FACS Canto II Flow cytometer and BD FACS Diva software, respectively. Data were entered and analysed using SPSS v 23.0. RESULTS: Over a period of 2 years, a total of 1,115 suspected patients were tested for acute leukaemia. Among them, 728 (65.3%) were males and 387 (34.7%) were females, with mean age 28 ± 21 years, ranging from 1 week to 87 years. Among a total of 875/1115 (78.5%) diagnosed cases of acute leukaemia, AML was the most common leukaemia present in 408/875 (46.6%) patients followed by B-ALL and T-ALL in 384/875 (43.8%) and 70/87 (8%) patients, respectively (p = 0.5712). Aberrant CD markers were detected in 109/875 (12.5%) leukaemias (p = 0.0628). The most common aberrant CD markers in B-ALL were CD13 and CD33 present in 30/384 (7.8%) cases separately. Among AML and T-ALL most common aberrant CD markers were CD7 and CD33 present in 25/408 (6.13%) and 7/70 (10%) cases, respectively. CONCLUSION: Special consideration should be given to the presence of aberrant CD markers when assigning lineages to acute leukaemias. They may be important diagnostic, prognostic, and management tools for institution of immunotherapy. KEY WORDS: Aberrant CD markers, Acute leukaemia, CD Markers, Flow cytometry, Immunophenotyping.

Diagnostic accuracy of plain computed tomography in detecting acute cerebral venous sinus thrombosis keeping magnetic resonance venography as gold standard.

OBJECTIVE: To determine the diagnostic accuracy of plain computed tomography using the ratio between hounsfield unit and haematocrit of cerebral venous sinuses in cases of acute cerebral venous sinus thrombosis taking magnetic resonance venography as the gold standard. METHODS: The cross-sectional validation study was conducted at the Department of Diagnostic Radiology, Combined Military Hospital, Rawalpindi, Pakistan, from March 9 to September 8, 2021, and comprised patients regardless of age and gender presenting with acute neurological and visual signs and symptoms of cerebral venous sinus thrombosis for <5 days. The patients were brain-imaged on 128-slice computed tomography scanner, and the image was assessed and the attenuation values in terms of Hounsfield unit of dural venous sinuses were calculated by taking appropriate region of interest. Haemoglobin and haematocrit values were noted from blood reports, and then the ratio between Hounsfield unit and haematocrit ratio was calculated. Magnetic resonance venography of the patients were performed and the patients were assessed for dural venous thrombosis. Data was analysed using SPSS 23. RESULTS: Of the 201 patients, 98(48.8%) were males and 103(51.2%) were female. The overall mean age was 35.32±19.707 years (range: 1 month-70 years). According to the Hounsfield unit-haematocrit ratio, acute cerebral venous sinus thrombosis was detected in 173(86.01%) patients, while magnetic resonance venography detected 178(88.6%). The Hounsfield unit-haematocrit ratio had sensitivity 91.01%, specificity 52.17% and diagnostic accuracy 86.57%. CONCLUSIONS: Computed tomography attenuation value and Hounsfield unit-haematocrit ratio on unenhanced computed tomography could be used as a reliable method to detect acute cerebral venous sinus thrombosis in emergency settings.

Development and Validation of In-house HEp-2 Cell Slides for Detection of Antinuclear Antibodies (ANA) by Indirect Immunofluorescence.

OBJECTIVE: To develop and validate in-house HEp-2 cell slides for the detection of ANA by indirect immunofluorescence. STUDY DESIGN: Cross sectional validation study. Place and Duration of the Study: Department of Immunology, Armed Forces Institute of Pathology Rawalpindi, Punjab, Pakistan, from April to September 2022. METHODOLOGY: This study involved development of in-house HEp-2 cell slides after procuring cell lines, sub-culturing and fixing them on different slides using variety of fixatives under different protocols. After standardisation of procedure, validation of procedure was done by testing sera of 305 patients for ANA detection at 1:40 dilution on in-house HEp-2 cell slides and subsequently on commercial HEp-2 cell slides (gold standard). Indirect immunofluorescence was observed by the two observers working independently and kept blinded from the results interpreted by each other. Data were collected on a pre-designed proforma and then analysed. Sensitivity, specificity, and positive predictive value (PPV) and negative predictive values (NPV) of in-house HEp-2 cell slides were calculated. RESULT: Sera of 305 patients were tested on in-house and commercial HEp-2 cell slides. Sensitivity and specificity of in-house HEp-2 cell slides for ANA detection were 96.92% and 99.58%, respectively. PPV and NPV of in-house HEp-2 cell slides came out to be 98.43% and 99.17% respectively. CONCLUSION: In-house HEp-2 cell slides are as effective as commercial HEp-2 cell slides for the detection of ANA and can be used as cost-effective alternative. KEY WORDS: Antinuclear antibodies (ANA), Human epithelial type-2 (HEp-2), Immunofluorescence.