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Bibliography

Clinical Method Validation

Human Blood Stabilisation - Immunophenotyping

2007

KEY PAPER: Advances in CD4 cell enumeration in resource-poor countries

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

Baum L, Crowe S. Landay AL

Publication

Current Opinion in HIV and AIDS (2007) 2, 157-245

The availability of antiretroviral medications increases the need for CD4 cell assays to monitor disease progression and the efficacy of therapy in resource-poor countries. Simplified flow cytometry is needed in centralized clinics, but there is a critical need for even less expensive easier methodology that can be used in smaller laboratories and in rural villages. TransFix can be used to stabilise samples for transportation to central laboratories.

http://www.ncbi.nlm.nih.gov/pubmed/19372892

2001

Affordable CD4+ T cell counts by flow cytometry. II. The use of fixed whole blood in resource poor settings

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

I. Jani, G.Janossy, A.Iqbal, F. Mhalu, E.Lyamuya, G.Biberfield, D.Glencross, L.Scott, J.T.Reilly, V.Granger & D.Barnett

Publication

Journal of Immunological Methods (2001) 257: 145-154

The feasibility and precision of CD4 counting in resource poor areas was tested using TransFix. It has been shown that immunological gating, including the primary identification of T, B and NK cells by virtue of their specific antibody binding capacity, is a robust method for absolute cell counting. With TransFixed samples primary CD4+ gating can be performed with great precision. There was also no difference in the precision of CD4+ counts performed on fresh cells in Tanzania and TransFixed cells in South Africa. TransFixed samples from smaller clinics can be batched over a period of one week for shipment to local or regional hospitals for analysis.

http://www.ncbi.nlm.nih.gov/pubmed/11687248

CD45 assisted panleucogating for accurate, cost effective dual platform CD4+ T cell enumeration

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

Deborah Glencross, Lesley E. Scott, Ilesh V. Jani, David Barnett and George Janossy

Publication

Cytometry (2001) 50: 69-77

A higher accuracy and precision of CD4 counts was documented with Pan Leucogating compared with lymphocyte gating using stabilised blood controls provided by UK NEQAS. These results were verified with 183 fresh samples and 112 TransFixed samples. These observations showed that dual platform leucocyte counts should replace lymphocyte counts.

http://onlinelibrary.wiley.com/doi/10.1002/cyto.10068/abstract

2015

Collection, Storage, and Preparation of Human Blood Cells

Sample Type: Monocytes, T lymphocytes, B lymphocytes, neutrophils, and platelets

Analysis Method: Flow Cytometry

Authors

P. K. Dagur, J. P. McCoy

Publication

Current Protocols in Cytometry 73 (5): 5.1-5.1.16

Human peripheral blood is often studied by flow cytometry in both the research and clinical laboratories. The methods for collection, storage, and preparation of peripheral blood will vary depending on the cell lineage to be examined as well as the type of assay to be performed. This unit presents protocols for collection of blood, separation of leukocytes from whole blood by lysis of erythrocytes, isolating mononuclear cells by density gradient separation, and assorted non-flow sorting methods, such as magnetic bead separations, for enriching specific cell populations, including monocytes, T lymphocytes, B lymphocytes, neutrophils, and platelets, prior to flow cytometric analysis. In some instances, such as shipping specimens from a distant site, it may be necessary to store blood for brief periods of time prior to staining. Several commercially available reagents, such as TransFix can be used to treat blood for storage. As an example of using these, a protocol for using Transfix is provided in this publication.

http://onlinelibrary.wiley.com/doi/10.1002/0471142956.cy0501s73/abstract

2002

Less expensive CD4+ T cell monitoring using panleucogating

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

Wilja M, Janossy G, Glencross D, Barnett D

Publication

Poster: Barcelona Aids Meeting - July 2002

Tests were carried out to compare the cost and accuracy of DP:lymphocyte gating using TriTEST with MultiSET software [LyG] and DP:panleucogating [PLG] and to evaluated Transfix to stabilize blood and extend the time between blood draw and testing. EDTA blood samples were analyzed within 24 hours by FACSCount, LyG and PLG. Transfix samples were evaluated after 3 and 7 days. Cells stabilized by Transfix, the correlation between results at day 3 and day 0 was excellent for PLG and FACSCount. Correlation between results at day 7 and day 0 were good only for PLG [R2=0.85]. In conclusion the PLG results correlated more closely than LyG with FACSCount results and Transfix allowed accurate testing for at least 3 days following blood draw.

http://www.iasociety.org/Abstracts/A5137.aspx

2003

Monitoring of human immunodeficiency virus infection in resource-constrained countries

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

S Crowe, S Turnbull and R Oelrichs

Publication

Clinical Infectious Diseases 37 (Supplement 1): S25-S35. 2003

The reference standards used to monitor human immunodeficiency virus (HIV) infection are flow cytometric analysis of T lymphocyte subsets to provide the CD4+ T cell count and molecular assays to quantify plasma HIV load. Few laboratories in resource-constrained countries can afford to perform these tests. This review discusses the above assays and their role in addition to clinical monitoring in resource-constrained countries. It states that TransFix TM stabilises whole blood for CD4 cell analysis for 7 days at 37 degrees Celsius and <3 days at 42 degrees Celsius. This stabilisation allows the transport of samples from distant sites to laboratories.

http://cid.oxfordjournals.org/content/37/Supplement_1/S25.short

2002

New concepts in affordable cd4+ t cell enumeration for resource-poor settings

Sample Type: CD4, CD8 and CD45+ cells

Analysis Method: Flow Cytometry

Authors

Janossy G, Jani IV, NJ Bradley, Pitfield T

Publication

International Conference on AIDS (2002) 7-12; 14: abstract no. MoPeB3220

Flow cytometers can provide accurate CD4+ T cell counts for monitoring HIV disease but remain too expensive for resource-poor settings. In this research they combined six modern concepts (i-vi) in flow cytometry (FCM) for more efficient and economical CD4 T cell enumeration. Volumetric flow cytometers were used with (ii) CD4, CD8 and CD45 generic monoclonal antibodies (iii) using the panleucogating protocol (iv) on TransFix stabilized blood samples. A simple protocol using generic MAb's, and with no hidden costs such as microbeads, allowed the accurate reporting of 16 parameters including leukocyte absolute counts and differentials, CD4 and CD8 absolute and percentage values, total CD4+CD8 T cell counts as well as CD4/CD8 ratios. Correlations for absolute CD4 counts and percentage values were R2>0.96, with virtually no bias. The correlation for CD8 counts was R2=0.997, with a bias of 5.2% in favour of the CD3+,CD8+ test but with no significant influence on CD4/CD8 ratios. One single assistant processed 400 samples per day, which extrapolates to a throughput of 100,000 CD4 tests per year.

http://www.iasociety.org/Abstracts/A2320.aspx

2005

KEY PAPER: Performance of aged, TransFix treated blood in the Guava EasyCD4 and EasyCD8 Assays

Sample Type: CD4+ and CD8+ T cells

Analysis Method: Guava EasyCD4 and EasyCD8 assays.

Authors

A Mergai et al

Publication

12th conference on retroviruses and opportunistic infection (2005) : Poster

Monitoring CD4+ and CD8+ T cell counts has become essential in monitoring healthy individuals and patients infected with HIV / AIDS. The study reported the compatibility of 15 day old TransFix treated blood with the Guava EasyCD4 and EasyCD8 assays. The staining pattern for TransFixed samples over 15 days was almost identical to the staining pattern at day 0. At 30 days the staining pattern had started to show signs of deterioration but accurate counts could still be obtained.

http://www.millipore.com/publications

Human Blood Stabilisation - Circulating Tumour Cells

2015

KEY PAPER: A Novel Strategy for Detection and Enumeration of Circulating Rare Cell Populations in Metastatic Cancer Patients Using Automated Microfluidic Filtration and Multiplex Immunoassay

Sample Type: Circulating rare cells (CTCs, CMCs, CECs, CSCs)

Analysis Method: Automated microfluidic filtration and multiplex immunoassay

Authors

M. J. M. Magbanua, M. Pugia, J. S. Lee, M. Jabon, V. Wang, M. Gubens, K. Marfurt, J. Pence, H. Sidhu, A. Uzgiris, H. S. Rugo, J. W. Park

Publication

PLoS ONE 10(10): e0141166

In this study, the performance of a novel approach for detection and enumeration of multiple rare cell populations was evaluated in the blood of metastatic breast and lung cancer patients using an automated microfluidic filtration and multiplex immunoassay strategy. Different circulating rare cell populations were detected and enumerated, including circulating tumour cells (CTCs), circulating mesenchymal cells (CMCs), circulating endothelial cells (CECs), and putative circulating stem cells (CSCs). Simultaneous assessment of CTCs, CMCs, CSCs and CECs may provide new tools to study mechanisms of disease progression and treatment response/resistance.
The highly controlled filtration process and the multi-step staining parameters were optimised to minimise the detection of false positives in healthy donor blood. Use of TransFix®, along with controlled shipping and storage conditions contributed to the high rate of reportable results (98%).
Blood was collected into tubes containing K3EDTA and 0.45mL Transfix® in customised TransFix/EDTA Vacuum Blood Collection Tubes (TVT-09-50-45).

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0141166

CSF Stabilisation

2012

Clinical relevance of flow cytometric immunophenotyping of the cerebrospinal fluid in patients with diffuse large B-cell lymphoma

Sample Type: CSF (large B cell lymphoma)

Analysis Method: Flow Cytometry

Authors

R. Alvarez, J. Dupuis, 1, A. Plonquet, C. Christov, C. Copie-Bergman, F. Hemery, I. Gaillard, T. El Gnaoui, F. Kuhnowski, M. Bedoui, K. Belhadj, P. Brugières and C. Haioun

Publication

Annals of Oncology, Volume 23, Issue 5 , pages 1274-1279, 2012

Central nervous system (CNS) relapse is an uncommon but dramatic complication of diffuse large B-cell lymphoma (DLBCL). Several studies have demonstrated the superiority of cerebrospinal fluid (CSF) flow cytometry (FCM), as compared with conventional cytology (CC), in detecting occult leptomeningeal disease. The clinical relevance of a positive FCM still has to be clarified. Makes reference to use of TransFix to stabilise and transport CSF samples.

http://annonc.oxfordjournals.org/content/23/5/1274.short

2010

Clinical significance of occult cerebrospinal fluid involvement assessed by flow cytometry in non-Hodgkin's lymphoma patients at high risk of central nervous system

Sample Type: CSF (non-Hodgkin's lymphoma)

Analysis Method: Flow Cytometry

Authors

Juan-Manuel Sancho, Alberto Orfao, Sandra Quijano, Olga García, Carlos Panizo, Elena Pérez-Ceballos, Guillermo Deben, Antonio Salar, Eva González-Barca, Natalia Alonso, Jose-Antonio García-Vela, Javier Capote, Francisco-Javier Peñalver, Mariano Provencio, Jesús Arias, Josefa Plaza, Dolores Caballero, Marta Morado, Evarist Feliu, Josep-Maria Ribera

Publication

European Journal of Haematology (2010) 85, Issue 4, 321–328

Quote: The specific techniques and procedures used for the analyses of CSF samples have been previously described in detail (21). CSF samples were analysed in parallel using CC in the hospital of origin and centrally by FCM at the Cytometry Service of the University of Salamanca (Spain) using a standardised 11-parameter FCM immunophenotypic assay of CSF samples stabilised with 0.2 mL of TransFix added at the hospital of origin.

http://www.ncbi.nlm.nih.gov/pubmed/20528905

2012

Current strategies in the diagnosis of diffuse large B-cell lymphoma of the central nervous system

Sample Type: CSF (large B cell lymphoma)

Analysis Method: Flow Cytometry

Authors

Alexander Baraniskin, Martina Deckert, Gernot Schulte-Altedorneburg, Uwe Schlegel, Roland Schroers

Publication

British Journal of Haematology, Volume 156, Issue 4, pages 421–432, February 2012

A detailed review of current strategies used in the diagnosis of diffuse large B cell lymphoma. The use of flow cytometry to detect CSF involvement in Primary CNS Lymphoma (PCNSL )provides improved diagnostic sensitivity as compared to cytopathology alone. With the availability of TransFix it is now possible to preserve CSF for up to 10 days. Accordingly it is now possible to analyse CSF samples from PCNSL patients in a central laboratory, preferably within a large multi-centre study.

http://onlinelibrary.wiley.com/doi/10.1111/bjh.2012.156.issue-4/issuetoc

2014

Detection & outcome of occult Leptomeningeal disease in diffuse large B-cell lymphoma and Burkitt lymphoma

Sample Type: CSF (diffuse large B-cell lymphoma and Burkitt lymphoma)

Analysis Method: Flow Cytometry

Authors

WH Wilson, JEC Bromberg, M Stetler-Stevenson

Publication

Haematologica. 2014 Jul;99(7):1228-35

Cytology of cerebrospinal fluid (CSF), the diagnostic gold standard has a low sensitivity with reported false negative rate of 20-60% suggesting that pre-treatment involvement of leptomeningeal disease is greater than initially reported. CSF was either analysed within an hour of lumbar puncture, or it was stabilised by two methods, one of which was using TransFix using the method outlined in the paper. Results provided the first evidence that occult CSF involvement by DLBCL or BL is clinically meaningful and associated with an adverse outcome. Results also indicated that patients at risk of CNS disease will benefit from intrathecal chemotherapy.

http://www.ncbi.nlm.nih.gov/pubmed/24727817

2015

Detection of central nervous system involvement in childhood acute lymphoblastic leukemia by cytomorphology and flow cytometry of the cerebrospinal fluid

Sample Type: CSF (lymphoblastic leukaemia)

Analysis Method: Flow Cytometry

Authors

S. Ranta, F. Nilsson, A. Harila-Saari, L. Saft

Publication

Pediatric Blood & Cancer (2015) 62(6): 951

The authors retrospectively compared flow cytometric immunophenotyping (CFI) of CSF with cytomorphology (CM) at diagnosis or relapse of childhood acute lymphoblastic leukaemia (ALL). They concluded that FCI of CSF increased the detection rate of CNS involvement of ALL approximately two times compared to cytomorphology, and that patients with low level CNS involvement may bene?t from additional intensi?ed systemic or CNS-directed therapy. CSF specimens were processed immediately on receipt in the laboratory or ?xed in Trans?x and processed the next morning. The authors note that one of the main problems in analysing CSF is the rapid cell death of leukocytes after sampling. Therefore, the FCI and CM results also always re?ect the time from sampling until the start of analysis. Routine use of stabilization media or ?xative immediately after sampling could further facilitate detection of malignant cells.

http://onlinelibrary.wiley.com/doi/10.1002/pbc.25363/full

2013

Detection of occult cerebrospinal fluid involvement during maintenance therapy identifies a group of children with acute lymphoblastic leukemia at high risk for relapse

Sample Type: Lymphoblasts in CSF (ALL)

Analysis Method: Flow Cytometry

Authors

C. Martínez-Laperche, A. M. Gómez-García, Á. Lassaletta, C. Moscardó, J. L. Vivanco, J. Molina, J. L. Fuster, J. M. Couselo, J. Sánchez de Toledo, E. Bureo, L. Madero, M. Ramírez

Publication

American Journal of Hematology, 88(5): 359–364, May 2013

To determine the clinical significance of the levels of lymphoblasts in the cerebrospinal fluid (CSF) of children diagnosed with acute lymphoblastic leukaemia (ALL).CSF samples from 136 patients were analysed by conventional cytology (CC) and flow cytometry (FCM) at the time of diagnosis and after intra-thecal therapy (IT). CSF samples for analysis by FCM were treated with TransFix at sample collection to facilitate transport of samples from 8 different hospitals to one centre for analysis. The results support previous findings that FCM analysis was more sensitive than CC in the detection of leukemic cells present in the CSF samples and can provide a powerful tool in detecting residual leukemic cells in the central nervous system.

http://onlinelibrary.wiley.com/doi/10.1002/ajh.23407/full

2015

Diagnostic and prognostic significance of flow cytometry immunophenotyping in patients with leptomeningeal carcinomatosis

Analysis Method: Flow Cytometry

Authors

D. Subira, M. Simo, J. Illian, C Serrano, S. Castanon, R. Gonzalo, J. J. Granizo, M Martinez-Garcia, M. Navarro, J. Pardo and J. Bruna

Publication

Clinical & Experimental Metastasis (2015) 32: 383-391

A multi-center study with eight patient recruitment centers throughout Spain that used TransFix CSF tubes to stabilise CSF samples from 166 patients. The study examined the sensitivity and specificity of flow cytometry (FC) diagnosis for leptomeningeal carcinomatosis. Compared with Cytology, FC showed greater sensitivity and negative predictive value, but lower specificity and positive predictive value. The multivariate analysis revealed that the percentage of CSF EpCAM positive cells predicted an increased risk of death.

http://www.ncbi.nlm.nih.gov/pubmed/25795393

2012

EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes

Sample Type: CSF and Vitreous biopsy

Analysis Method: Flow Cytometry

Authors

J. J. M. van Dongen, L. Lhermitte, S. Böttcher, J. Almeida, V. H. J. van der Velden, J. Flores-Montero, A. Rawstron, V. Asnafi, Q. Lécrevisse, P. Lucio, E. Mejstrikova, T. Szczepański, T Kalina, R. de Tute, M. Brüggemann, L Sedek, M. Cullen, A. W. Langerak, A. Mendonça, E. Macintyre, M. Martin-Ayuso, O. Hrusak, M. B. Vidriales, A. Orfao

Publication

Leukemia (2012) 26: 1908?1975

Section 3 discusses the design and evaluation of the 8-colour, 13 parameter Euroflow™ small sample tube (SST) and related sample preparation protocols aimed to evaluate CSF and Vitreous biopsy samples, in patients suspected of leukaemia. Samples collected either with culture medium with FBS of FCS, or in TransFix Sample Storage Tubes, which the author states in the general discussion ?result in optimal stabilization of the cells for a few days (for example, 48h) and improve cell detection?.

https://www.nature.com/leu/journal/v26/n9/pdf/leu2012120a.pdf

2008

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

2012

Flow cytometry as a diagnostic tool in lymphomatous or leukemic meningitis

Sample Type: CSF (NM)

Analysis Method: Flow Cytometry

Authors

M.S. Ahluwalia, P.K. Wallace and D.M. Peereboom

Publication

Cancer, Volume 118, Issue 7, pages 1747 – 1753, 2012

In patients with neoplastic meningitis (NM), early diagnosis is highly desirable because the rapid institution of intrathecal therapy may mitigate the course of the disease. Cytology, long considered the "gold standard" for diagnosis, has low sensitivity because of both the paucity of cells in the cerebrospinal fluid (CSF) and morphological similarities between benign and malignant cells. A comprehensive review of the literature from 2005 through 2011 was performed that focused on diagnostic modalities for lymphomatous meningitis. Several studies demonstrated the sensitivity of flow cytometry to be several-fold higher than that of cytology for the detection of CSF leukemia/lymphoma. Makes reference to use of TransFix to stabilise and transport CSF samples.

http://onlinelibrary.wiley.com/doi/10.1002/cncr.26335/full

2017

Guidelines For Diagnosis, Prevention And Management Of Central Nervous System Involvement In Diffuse Large B-Cell Lymphoma Patients By The Spanish Lymphoma Group (GELTAMO)

Sample Type: CSF (Diffuse Large B-Cell Lymphoma)

Analysis Method: Flow Cytometry

Authors

F. J. PeÑalver, J. M. Sancho, A. de la Fuente, M. T. Olave, A. Martín, C. Panizo, E. Pérez, A. Salar, A. Orfao

Publication

Haematologica (2017) 102: 235-245

This report by the Spanish Lymphoma Group (GELTAMO) aims to provide useful guidelines and recommendations for the prevention, diagnosis, and treatment of central nervous system diffuse large B-cell lymphoma patients with, or at risk of, leptomeningeal and/or brain parenchyma lymphoma relapse. The Authors summarise and recommend the use of standardised and validated >8-colour FCM evaluation of stabilised CSF in the diagnosis work-up of DLBCL patients at risk of CNS disease for the identification of occult CNSL, and state that immediate sample preservation (preferably in TransFix) is strongly recommended.

http://www.haematologica.org/content/102/2/235.full.pdf+html

2014

KEY PAPER: Guidelines on the use of multicolor flow cytometry in the diagnosis of haematological neoplasms

Analysis Method: Flow Cytometry

Authors

Ulrika Johansson, David Bloxham, Stephen Couzens, Jennifer Jesson, Ricardo Morilla, Wendy Erber, Marion Macey and British Committee for Standards in Haematology

Publication

British Journal of Haematology, Volume 165, Issue 4, pages 455–488, May 2014

These guidelines have been developed by UK-based clinical flow cytometry (FC) experts along with clinical representatives to take into account advances in the state of the art of clinical flow cytometry since 2002. Their work was revised by the Haemato-oncology Task Force of the British Committee for Standards in Haematology (BCSH) before review by other UK-based Haematologists and FC experts, the BCSH and the British Society for Haematology committee. This document covers a varied range of topics, from selection and setup of flow cytometers through data analysis to training considerations for staff working in the clinical FC laboratory.The guidelines strongly recommend sample age and quality is assessed prior to analysis and in the case of cerebrospinal fluid (CSF), it is recommended that unless immediate analysis is possible then samples can be stored in TransFix/EDTA CSF Sample Storage Tubes for up to 72 hours before analysis*. *TransFix/EDTA CSF Sample Storage Tubes are currently a Research Use Only product and Cytomark recommends that internal validation of the product must be completed by the end user before considering use of the products in a clinical setting.

http://onlinelibrary.wiley.com/doi/10.1111/bjh.12789/abstract

2017

Higher rate of central nervous system involvement by flow cytometry than morphology in acute lymphoblastic leukemia

Sample Type: CSF (ALL)

Analysis Method: Flow Cytometry

Authors

J. Dass, A. Dayama, P. C. Mishra, M. Mahapatra, T. Seth, S. Tyagi, H. P. Pati, R. Saxena

Publication

Int J Lab Hem.?39: 546-551.?https://doi.org/10.1111/ijlh.12694

This paper investigates the use of flow cytometry (FCM) as apposed to traditional cytopathology methods for diagnosis of central nervous system (CNS) involvement in acute lymphoblastic leukaemia (ALL). In the discussion, the author discusses the limitations of FCM, such as the viability declines rapidly in cerebrospinal fluid (CSF) samples, and cell degeneration occurs within 1 hour of sample collection. As a solution, the author recommends the use of TransFix to stabilise CSF for up to 72 hrs.

http://onlinelibrary.wiley.com/doi/10.1111/ijlh.12694/full

2016

How to facilitate early diagnosis of CNS involvement in malignant lymphoma

Sample Type: CSF (malignant lymphoma)

Analysis Method: Flow Cytometry

Authors

A. Korfel, M. Nowosielski, J. Pardo-Moreno, F. J. Penalver, G. Buda, H. Bennani, M. Costopoulos, M. Le Garff-Tavernier, C. Soussain, M. Schmid, J. A. Orfao, M. Glantz

Publication

Expert Review of Hematology, 9 (11) https://doi.org/10.1080/17474086.2016.1242405

The authors discuss several analysis methods to facilitate early diagnosis of CNS involvement in malignant lymphoma (CD19/10/45/kappa/lambda for B cell Lymphomas). They recommend stabilising CSF with TransFix if samples cannot be analysed within 1 hour for flow cytometric analysis.

http://www.tandfonline.com/doi/abs/10.1080/17474086.2016.1242405

2009

Identification of Leptomeningeal Disease in Aggressive B-Cell Non-Hodgkin's Lymphoma: Improved Sensitivity of Flow Cytometry

Sample Type: CSF neoplastic cells from B-NHL

Analysis Method: Flow Cytometry

Authors

Quijano S., López A, Sancho JM, Panizo C, Debén G, Castilla C, García-Vela JA, Salar A, Alonso-Vence N, González-Barca J, Peñalver FJ, Plaza-Villa J,Morado M, José García-Marco, Jesús Arias, Javier Briones, Secundino Ferrer, Javier Capote, Concepción Nicolás, and Alberto Orfao

Publication

Journal of Clinical Oncology (2009) 27: 1462 - 1469.

We evaluate the sensitivity and specificity of a new 11-parameter Flow cytometry (FCM) approach versus conventional cytology (CC) for detecting neoplastic cells in stabilized CSF samples from newly diagnosed aggressive B-cell non-Hodgkin’s lymphoma (B-NHL) at high risk of CNS relapse, using a prospective, multicentric study design.

For multi-parameter FCM analyses, CSF samples (median volume, 2.0mL; range, 0.5 to 4.0 mL) were directly collected into tubes containing EDTA and 0.2 mL of TransFix (Cytomark) and shipped overnight to the central FCM laboratory. Immediately on arrival at the central laboratory, the volume of the CSF sample was measured (after subtracting 0.2 mL corresponding to the TransFix solution) and recorded;

http://www.ncbi.nlm.nih.gov/pubmed/19224854

2006

Identification of Neoplastic Infiltration of the Cerebrospinal Fluid (CSF) in Patients with Aggressive B-Cell Non-Hodgkin’s Lymphoma (B-NHL) without Clinical Evidence of Leptomeningeal Disease: A Comparative Analysis of the Utility of Flow Cytometry (FCM) Versus Conventional Cytology (CC)

Sample Type: CSF (neoplastic cells, B-NHL)

Analysis Method: Flow Cytometry

Authors

Orfao A, Quijano S, Lopez A, Deben G, Salar A, Poderos C, Sancho JM, Vallejo C, Arias J, Garcia JA, Capote F, Morado M, Nicolas C, Briones J, Fernandez S,Carmona, L. Vazquez, and J.F. San Miguel

Publication

Blood (ASH Annual Meeting Abstracts) (2006) 108: 4605.

Quote: In all cases, the CSF samples were analysed simultaneously by CC at the institution of origin and FCM, centrally one institution. For the FCM analysis of the CSF, stabilised samples (TransFix, CYTOMARK) were systematically stained with the following combination of monoclonal antibodies: CD8-sIgl/CD56-sIgk/CD4-CD19/CD3/CD20/CD45 (FITC/PE/PERCPCY5.5/PECY7/APC/APCCY7). If the FCM test showed infiltration, an additional 6-color antibody panel was used for full phenotypic characterisation of the disease.

http://abstracts.hematologylibrary.org/cgi/content/short/108/11/4605

2016

Leukemic blasts are present at low levels in spinal fluid in one-third of childhood acute lymphoblastic leukemia cases

Sample Type: CSF (B-cell precursor ALL and T-cell ALL)

Analysis Method: Flow Cytometry

Authors

M. Levinsen, H. V. Marquart, L. Groth-Pedersen, J. Abrahamsson, B. K. Albertsen, M. K. Andersen, T. L. Frandsen, A. Harila-Saari, C. Pronk, A. Ulvmoen, G. Vaitkevi?ien?, P. M. L?hteenm?ki, R. Niinim?ki, M. Taskinen, M. Jeppesen, K. Schmiegelow

Publication

Pediatr Blood Cancer, 63 (11): 1935-1942

The authors assessed centralised ?ow cytometry of locally TransFix-?xed cerebrospinal ?uid (CSF) samples versus local conventional cytospin-based cytology for detecting leukemic cells and evaluating kinetics of elimination of leukemic cells in CSF. CSF samples were collected into tubes containing TransFix to stabilise B-cell precursor ALL (CD3/CD10/CD19/CD20/CD34/CD38/CD45), and T-cell ALL (CD3/CD4/CD7/CD8/CD45/CD56), allowing samples to be shipped for centralised analysis, preferably within three days of lumbar puncture. To verify leukemic classification, the CSF from 2 patients were flow sorted followed by FISH targeting the leukemic karyotypes. The authors recommend the use of on-site fixation and rapid centralised flow cytometric analysis of CSF samples.

http://onlinelibrary.wiley.com/doi/10.1002/pbc.26128/full

2012

Role of flow cytometry immunophenotyping in the diagnosis of leptomeningeal carcinomatosis

Sample Type: CSF (leptomeningeal carcinomatosis)

Analysis Method: Flow Cytometry

Authors

D. Subirá, C. Serrano, S. Castañón, R. Gonzalo, J. Illán, J. Pardo, M. Martínez-García, E. Millastre, F. Aparisi, M. Navarro, M. Dómine, I. Gil-Bazo, P. Pérez Segura, M. Gil and J. Bruna

Publication

Neuro-Oncology, Volume 14, Issue 1, pages 43-52, 2012

Purpose: To explore the contribution of flow cytometry immunophenotyping (FCI) in detecting leptomeningeal disease in patients with solid tumors. CSF samples obtained from a lumbar puncture (n = 77) and from an Ommaya reservoir (n = 1) were collected in EDTA tubes containing 0.2 mL of an immunofixative reagent (TransFix, Cytomark), necessary to guarantee safe transportation. FCI seems to be a promising new tool for improving the diagnostic examination of patients with suspicion of LC. Detection of epithelial cells with a higher DNA content is highly specific of LC, but evaluation of the nonepithelial cell compartment of the CSF might also be useful for supporting this diagnosis.

http://neuro-oncology.oxfordjournals.org/content/14/1/43.short

2017

KEY PAPER: Study of body fluid samples using flow cytometry: Six years of experience at the Hospital Universitario San Ignacio -Ponti cia Universidad Javeriana, Bogota - Colombia

Sample Type: Cerebrospinal fluid, bronchoalveolar lavage, pleural fluid, pericardial fluid, ascite fluid

Analysis Method: Flow Cytometry

Authors

A. Campos, L. Trujillo, D. López, L. Beltrán, E. Arias, G. Vélez, A. Infante, I. De Los Reyes, M. Vizcaíno, P. C. Guzmán, M. V. Herrera, J. Solano, D. Londo?o, A. Ca?as, F. Pretelt, J. C. Pérez, C. Cardozo, S. Fiorentino, S. Quijano

Publication

Universitas Scientiarum, 22(2): 123 - 143, 2017. doi: 10.11144/Javeriana.SC22-2.sobf

The Authors discuss findings from 6 years of experience in implementing flow cytometry in Columbia. The authors used TransFix to stabilise all sample, consisting of mainly cerebrospinal fluid, but also bronchoalveolar lavage, pleural fluid, pericardial fluid and ascite fluid from patients with acute and chronic leukaemia, myelodysplastic syndromes, lymphomas, myeloma, autoimmune diseases, immunodeficiencies and solid tumours. The Authors final conclude that this work represents the first report at the national level supporting Transfix implementation in pre-analytical FCM studies of all body fluids that are processed in the clinical practice.

https://search.proquest.com/openview/538464d63f7cc2192885d3559b2871f6/1?pq-origsite=gscholar&cbl=2041198

Ten-color 15-antibody flow cytometry panel for immunophenotyping of lymphocyte population

Sample Type: CSF lymphocytes

Analysis Method: Flow Cytometry

Authors

A. Rajab, O. Axler, J. Leung, M. Wozniak, A. Porwit

Publication

International Society for Laboratory Hematology (2017) 39 (S1): 76-85

The Authors of this paper have developed a lymphoproliferative disorder screening tube (LPD- ST) with the aim to provide comprehensive immunophenotyping of lymphocyte subsets with minimal need for additional testing. The LPD- ST consists of CD4/kappa FITC, CD8/lambda PE, CD3/CD14ECD, CD38PC5.5, CD20/CD56PC7, CD10APC, CD19APC- A700, CD5APC- A750, CD57/CD23PB and CD45KO. They also claim that this new LPD- ST is more suitable with paucicellular samples such as cerebrospinal fluid (CSF) than previous lymphocyte subset panels. In their experiments, the Authors fix CSF samples in TransFix, and it is inferred that flow results match immunohistochemical staining results.

http://onlinelibrary.wiley.com/doi/10.1111/ijlh.12678/full

Bone Marrow Stabilisation

2016

The utilisation of modern transport and telecommunications platforms to assist in the remote provision of paediatric cancer diagnostics in Tanzania

Sample Type: Bone marrow

Analysis Method: Flow Cytometry

Authors

M. Mcdermott, K. O'Hare, P. Scanlon, D. Grehan, S. Rooney, R. Laiti, J. Kaijage, S. Jumanne, M. O'Sullivan

Publication

Pediatric Blood and Cancer, 63: Supplement 3 (S71)

Conference Abstract: Interim diagnostic services were provided by OLCHC in Dublin for paediatric cancer screening in Tanzania utilising DHL to transport fixed bone marrow samples and communicating the results via the WhatsApp communication platform in an average of 2.9 days. The bone marrow samples were fixed with TransFix to maintain integrity during transportation.

https://www.cytomark.co.uk/downloads/Mcdermott_2016_(conference_abstract).pdf


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