FLOWCYTOMETRY TESTS

FLOWCYTOMETRY (IMMUNOPHENOTYPING)

FLOWCYTOMETRY (IMMUNOPHENOTYPING)
B5073	Immunophenotyping of Leukemia BY FLOWCYTOMETRY Selective panels of antibodies are used for cell surface/intracellular markers to evaluate and diagnose specific Acute Leukemia types based on the clinical diagnosis and other primary lab findings. Report interpreted based on the percentage of expression of CD markers. Panel used: A tube with CD45 for screening blasts as well as to detect background fluorescence and autoflourescence. This will be followed by a three tube 8 colour primary leukemia panel. Additional secondary tubes will be processed for T ALL, AML-M6 & M7 and other scenarios based on the requirements (B5076 – Leukemia secondary panel)	5-6 ml of Sodium heparin (Green top) anti-coagulated peripheral blood, bone marrow samples. 2-3 ml of other body fluids pericardial fluid, ascitic fluid, CSF, pleural fluid. Storage: Bone marrow and peripheral blood samples will be stored at room temperature for few hours or stored at 4°C for next day processing. CSF and fluids will be freshly processed.
B5073	Immunophenotyping of Lymphoma BY FLOWCYTOMETRY Selective panels of antibodies are used for cell surface markers to evaluate and diagnose specific Lymphomas based on the clinical diagnosis and other primary lab findings. Panel used: A primary screening tube with CD45, CD3, CD7, CD19, CD4, CD8 along with surface kappa and lambda for primary lymphoma screening. This will be followed by secondary B NHL/CLPD tubes (2 tubes 8 colour), Hairy cell Leukemia tube (single tube 8 colour) or T/NK secondary tubes (2 tubes 8 colour) based on the requirements.	5-6 ml of Sodium heparin (Green top) anti-coagulated peripheral blood, bone marrow samples. 2-3 ml of other body fluids pericardial fluid, ascitic fluid, CSF, pleural fluid. Storage: Bone marrow and peripheral blood samples will be stored at room temperature for few hours or stored at 4°C for next day processing. CSF and fluids will be freshly processed.
B5074	Immunophenotyping of pid/ alps/ LYMPHOCYTE SUBSET ANALYSIS BY FLOWCYTOMETRY Selective panels of antibodies are used for cell surface markers to evaluate and diagnose primary immunodeficiency disorders based on the clinical diagnosis and other primary lab findings and also to screen for immune reconstitution post-transplant. Report interpreted as absolute counts based on the percentage of expression of CD markers. Panel used: A single tube, 8 colour, 12 antibody panel (PIDOT tube recommended by EUROFLOW for PID screening).	5-6 ml of Sodium heparin anti-coagulated (Green top) peripheral blood sample of the patient. Stored at 4°C.
B5074	IMMUNOPHENOTYPING FOR LAD1 (Leukocyte Adhesion Deficiency) BY FLOWCYTOMETRY Selective panels of antibodies are used for cell surface markers to evaluate and diagnose LAD based on the clinical diagnosis and other primary lab findings. Markers used for LAD Diagnosis: CD18, CD11a, CD11b, CD11c on patient and healthy control Gating strategy: FSC/SSC on granulocytes	5-6 ml of Sodium Heparin anti-coagulated (Green top) peripheral blood samples of a patient and healthy control. Stored at 4°C.
B5074	Dhr test for chronic granulomatous disease (cgd) Patient’s peripheral blood neutrophils are stimulated with a specific mitogen like PMA and the difference in fluorescence is used to diagnose CGD by comparing with a normal control
B5074	PLATELET IMMUNOPHENOTYPING FOR GLANZMANN'S THROMBASTHENIA (GT) AND BERNARD SOULIER SYNDROME (BSS) BY FLOW CYTOMETRY. Selective panel of cell surface antibodies used to diagnose GT and BSS, based on the clinical and other laboratory findings. Markers used for GT Diagnosis: CD42a (GPIb /IX) and CD41a (GPIIb) and CD61 (GPIIIa) on healthy control and patient. Gating strategy: FSC/SSC on platelets	5-6 ml of Sodium heparin anti-coagulated (Green top) peripheral blood samples of a patient and healthy control. Stored at 4°C.
B5075	Flow cytometry for Paroxysmal Nocturnal Hemoglobinuria (PNH) Selective panels of antibodies are used for cell surface markers to evaluate and diagnose PNH based on the clinical diagnosis and other primary lab findings. The panel is based on the 2018 ICCS consensus for PNH Markers used for PNH Diagnosis: FLAER (Alexa Fluor 488nm), CD59, CD15, CD64, CD14, CD45, CD24 on patient and healthy control Gating strategy: CD24 and FLAER expression on patient neutrophils (gated with CD15), CD14 and FLAER on monocytes (gated with CD64) and CD59 on RBCs (FSC/SSC, CD45/SSC, CD64/SSC and CD15/SSC).	5-6 ml of Sodium heparin anti-coagulated (Green top) peripheral blood samples of a patient and healthy control. Stored at 4°C.
B5076	lEUKEMIA SECONDARY PANEL Additional secondary tubes will be processed for T ALL, AML-M6 & M7 and other scenarios based on the requirements	Same sample used for the primary panel. No new sample required except in special scenarios (inadequate sample, viability issues)
B5081	CD34+ cells Enumeration Enumeration of CD34 positive cells in PBSCT and BMT samples Gating Strategy: ISHAGE gating strategy – CD34 kit based enumeration	1 ml of PBSC (or) bone marrow/5-6 ml of peripheral blood in sodium heparin anticoagulated (green top) tubes. Processed on the same day within 4 hours from the receiving time
B5081	DLI - CD3 Enumeration Enumeration of CD3 positive cells in PDLI sample to be given to the transplant recipient. Gating Strategy: ISHAGE gating strategy.	2 ml of apheresis / 5-6 ml of peripheral blood in sodium heparin anticoagulated (green top) tubes. Processed on the same day
MINIMAL RESIDUAL DISEASE (MRD) FOR HEMATOLOGICAL MALIGNANCIES
B5080	Minimal Residual Disease detection (MRD) for B cell ALL Selective panels of antibodies are used for cell surface markers to evaluate minimal residual disease by utilizing an LAIP based DFN approach. Each sample is processed in two tubes and 1.7 million events are acquired per tube. Panel and Markers Used for MRD of B CELL ALL: Two tube 8 colour panel. CD20, CD10, CD19, CD45, CD34, CD58, CD38, CD81, CD123, CD200, CD66c, CD73	5-6 ml of Sodium heparin (Green top) / EDTA anti-coagulated bone marrow samples. Storage: Bone marrow samples will be stored at room temperature for few hours or stored at 4°C for next day processing.
B5096	Minimal Residual Disease detection (MRD) for MULTIPLE MYELOMA Selective panels of antibodies are used for cell surface and cytoplasmic markers to evaluate minimal residual disease by utilizing the immunophenotypic difference between normal and clonal plasma cells. Each sample is processed in two tubes and 1.7 million events are acquired per tube. Panel and Markers Used for MRD of Multiple Myeloma: Two tube 8 colour panel. CD38, CD138, CD19, CD45, CD27, CD81, CD56, CD117, cytokappa and cytolambda	5-6 ml of Sodium heparin (Green top) anti-coagulated bone marrow samples. Myeloma samples for MRD should be processed on the same day.
B5109	Minimal Residual Disease detection (MRD) for T cell ALL Selective panels of antibodies are used for cell surface and cytoplasmic markers to evaluate minimal residual disease. Each sample is processed in two tubes and 1.7 million events are acquired per tube. Panel and Markers Used for MRD of T CELL ALL: CD3, cytoCD3, CD7, CD45, CD34, CD56, CD48, CD99, CD5, CD4, CD8	5-6 ml of Sodium heparin (Green top) / EDTA anti-coagulated bone marrow samples. Storage: Bone marrow samples will be stored at room temperature for few hours or stored at 4°C for next day processing.
B5110	Minimal Residual Disease detection (MRD) for AML Selective panels of antibodies are used for cell surface markers to evaluate minimal residual disease by utilizing an LAIP based DFN approach. Each sample is processed in two tubes with 10 antibodies and 1.7 million events are acquired per tube. Panel and Markers Used for MRD of AML: CD45, CD34, CD117, HLADR, CD64, CD14, CD33, CD13, CD11b, CD7, CD19, CD56, CD38, CD123, CD36, CD15.	5-6 ml of Sodium heparin (Green top) / EDTA anti-coagulated bone marrow samples. Storage: Bone marrow samples will be stored at room temperature for few hours or stored at 4°C for next day processing.

Apart from these routine tests, flow cytometry lab also provides immunophenotyping analysis as a part of specific transplantation-related graft manipulation or cell therapy procedures like TCRαβ depletion, CD45RA depletion, CD34 enrichment, CD56 enrichment etc.