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2008

Human Blood Stabilisation - Immunophenotyping

Research needs and challenges in the development of HIV diagnostic and treatment monitoring tests for use in resource-limited settings

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

B. Cheng, A. Landay, V. Miller

Publication

Current Opinion in HIV & AIDS (2008) 3: 495-503

The aim of this article is to review research priorities for current and new technologies to diagnose HIV and to monitor treatment response, including technologies to enumerate CD4 cell counts in resource-limited settings.
The Authors note that blood preservatives may allow for the preservation of whole blood so that the sample can be shipped to a centralized laboratory with up to 15 days (TransFix; UK NEQAS, Shef?eld, South Yorkshire, UK) from collection to testing. Blood preservatives may play a very important role in improving access to CD4 cell count testing as they will allow the sample to be transported to a laboratory where the testing will be performed.

http://journals.lww.com/co-hivandaids/Fulltext/2008/07000/Research_needs_and_challenges_in_the_development.14.aspx

CSF Stabilisation

Flow Cytometric Immunophenotyping of Cerebrospinal Fluid

Sample Type: CSF (neoplastic cells, B and T cell lymphomas and acute leukemias)

Analysis Method: Flow Cytometry

Authors

Jaco Kraan, Jan W. Gratama, Corinne Haioun, Alberto Orfao, Anne Plonquet, Anna Porwit, Sandra Quijano, Maryalice Stetler-Stevenson, Dolores Subira, Wyndham Wilson

Publication

Current Protocols in Cytometry - Published Online: 1 July 2008

Quote: Leptomeningeal disease is an important adverse complication occurring in patients with B and T cell lymphomas and acute leukemias of lymphoid and myeloid origin. Recent reports suggest that multiparameter flow cytometry immunophenotypic assessment of spinal fluid samples could improve the efficiency of detection of CNS involvement, due to its high specificity and greater sensitivity. However, spinal fluid samples are frequently paucicellular with a rapidly decreasing cell viability. Staining of spinal fluid therefore requires dedicated sample storage/transport, staining, and preparation protocols. The Basic Protocol in this unit outlines a consensus multiparameter (3- to 8-color) flow cytometry immunophenotypic protocol for the evaluation of CNS involvement of cerebrospinal fluid (CSF) samples by neoplastic cells. A Support Protocol describing the simultaneous assessment of surface and cytoplasmic antigens is also provided. Finally, in the Alternate Protocol, we describe a method to calculate absolute numbers of both normal and pathological cell subpopulations by adding counting beads to the assay.

http://onlinelibrary.wiley.com/doi/10.1002/0471142956.cy0625s45/abstract

Bone Marrow Stabilisation

Standardization of Flow Cytometric Minimal Residual Disease Evaluation in Acute Lymphoblastic Leukemia: Multicentric Assessment Is Feasible

Sample Type: Bone Marrow (BCP- and T-ALLs)

Analysis Method: Flow Cytometry

Authors

Dworzak, M. N., Gaipa, G., Ratei, R., Veltroni, M., Schumich, A., Maglia, O., Karawajew, L., Benetello, A., Pötschger, U., Husak, Z., Gadner, H., Biondi, A., Ludwig, W.-D. and Basso, G.

Publication

Cytometry B Clin Cytom. 2008 Nov;74(6):331-40.

Flow cytometric (FCM) assessment of minimal residual disease (MRD) in acute lymphoblastic leukaemia (ALL) patients was assessed at 3 different sites in Italy and one in Germany to see whether standardization was possible.MRD-evaluation by FCM in ALL can be standardized for reliable multicentric assessment in large trials.In order to assess the inter-laboratory concordance, artificial samples were created using TransFix. TransFix was used to stabilise leukemic cells from nine patients (seven B cell ALL and two T cell ALL) and these cells were mixed into bone marrow preparations at different concentrations. These were then sent to the sites for FCM analysis. The concordance between all four centres was 98%. Therefore, TransFix was shown to be successful in stabilising leukemic cells.

http://www.ncbi.nlm.nih.gov/pubmed/18548617


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