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Bibliography

Bibliography

Quality Assurance

Human Blood Stabilisation - Immunophenotyping

2007

A proficiency testing program for flow cytometry: Istanbul experience

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

Demirel GY, Topbas F

Publication

The first International Proficiency Testing Conference, Romania: Poster

TransFix was used to prepare External Quality Assurance samples for CD4 Immune Monitoring

2002

KEY PAPER: Evaluation of stabilised blood cell products as candidate preparations for quality assessment programs for CD4 T-cell counting

Sample Type: T-cell subsets

Analysis Method: Flow Cytometry

Authors

M. Bergeron, A.Shafaie, T.Ding, S.Phaneuf, N.Soucy, F. Mandy, J.Bradley & J.Fahey

Publication

Cytometry (2002) 50, 86-91

Exceptionally robust cell preparations are needed for quality assessment programs. A suitable product must withstand the environmental stress related to transportation for a minimum of 6 day. Six preparations were evaluated by monitoring T-cell subset values for samples stored at 4 degrees Celsius, 22 degrees Celsius and 37 degrees Celsius. Sample stability was tested daily up to day 7. Only TransFix and ImmunoTrol were successful at stabilising blood samples across the full range of storage temperatures.

http://www.ncbi.nlm.nih.gov/pubmed/12116350

2007

Flow cytometric CD34+ stem cell enumeration: Lessons from nine years' external quality assessment within the Benelux countries

Sample Type: CD34+ stem cell

Analysis Method: Flow Cytometry

Authors

W. H. B. M. Levering, F. W. M. B. Preijers, W. N. van Wieringen, J. Kraan, W. A. M. van Beers, K. Sintnicolaas, D. J. van Rhenen, J. W.Gratama

Publication

Cytometry 72B: 178-188 doi:10.1002/cyto.b.20351

Assessment of interlab variability with regard to Flow Cytometric CD34+ Stem Cell enumeration. Authors discuss the use of TransFix in an EQA scheme to improve interlaboratory variability.

http://onlinelibrary.wiley.com/doi/10.1002/cyto.b.20351/full

2010

Interlaboratory variability of CD34+ stem cell enumeration. A pilot study to national external quality assessment within the Czech Republic

Sample Type: CD34+ stem cells

Analysis Method: Flow Cytometry

Authors

D. LYSÁK, T. KALINA, J. MARTÍNEK, Z. PIKALOVÁ, D. VOKURKOVÁ, M. JAREŠOVÁ, I. MARINOV, A. ONDREJKOVÁ, M. ŠPACEK, O. STEHLÍKOVÁ

Publication

International Journal of Laboratory Hematology (2010) 32, e229-e236

Quote: The study took place between November 2007 and May 2009 and consisted of three surveys. Ten laboratories took part in the study, but only nine laboratories participated in the first survey. Each laboratory was assigned a unique number (ULN) to retain confidentiality. Two samples of mobilized peripheral blood were included in every survey. Samples were stabilized with TransFix (Cytomark Ltd, Buckingham, UK),divided in 1 ml aliquots and shipped overnight by courier to the participants. The blood was collected from patients or healthy donors who underwent stem cell mobilization after giving informed consent. All subjects were negative for infectious diseases (humanimmunodeficiency virus, hepatitis B and C, syphilis).The stability of the samples was assured for at least 3 days (CV up to 5% in the stability study).Each laboratory performed the CD34+ enumeration according to its local protocol and practice and reported the measured values (leukocyte count,CD34+ percentage and absolute count) and some methodological details. All data were analyzed in the co-ordinating laboratory, and the results sent to the participants. The absolute CD34+ count was assignedas an indicator of laboratory performance.

http://www.ncbi.nlm.nih.gov/pubmed/20561093

2003

New trends in affordable CD4+ T-cell enumeration by flow cytometry in HIV/AIDS

Sample Type: CD4+ cells

Analysis Method: Flow Cytometry

Authors

Janossy G, Jania IV, Brandob B

Publication

Clinical & Applied Immunology (2003) 4: 91-107

Inexpensive Antiretroviral Therapy (ART) might soon be available to treat human immunodeficiency Virus (HIV) infections in resource-restricted areas of the globe. The number of CD4+ T-cells in the blood is the single most important laboratory parameter to select patients for therapy at the right time and to monitor the effect of ART. Stabilised blood cell standards and blood stabilising fluid, referred to as TransFix have recently been introduced for Quality Assurance. As the fast changing scenery of Flow Cytometry diminished the influence of company-orientated and instrument-orientated Quality Assurance Schemes, more emphasis had to be paid to the use of stabilised standard biological samples in international Quality Assurance Schemes. Preservative fluids, such as TransFix have been introduced with two beneficial effects. Firstly, TransFix added to whole blood at 1:10 dilution stabilises whole blood to perform reliable lymphocyte subset analysis for as long at 10 days. The catchment areas for laboratories can therefore be extended to receive samples from further afield. Secondly. TransFix has an additional bonus for quality assurance and interlaboratory technical standardisation. Samples shipped and tested within the time limits of 10 days between continents can be fully utilised to check the performance of instruments, reagents, full technologies and other parameters.

http://www.sciencedirect.com/science/article/pii/S1529104903000497

Human Blood Stabilisation - Other

2015

Quantification of Cells with Specific Phenotypes I: Determination of CD4+ Cell Count Per Microliter in Reconstituted Lyophilised Human PBMC Prelabeled with Anti-CD4 FITC Antibody

Sample Type: Human peripheral blood mononuclear cells

Analysis Method: Flow Cytometry

Authors

Stebbings R, Wang L, Sutherland J, Kammel M, Gaigalas AK, John M, Roemer B, Kuhne M, Schneider RJ, Braun M, Engel A, Dikshit DK, Abbasi F, Marti GE, Sassi MP, Revel L, Kim SK, Baradez MO, Lekishvili T, Marshall D, Whitby L, Jing W, Ost V, Vonsky M, Neukammer J.

Publication

Cytometry Part A (2015): 87A: 244-253

A surface-labelled lyophilised lymphocyte (Sll) preparation has been developed using human peripheral blood mononuclear cells prelabelled with a fluorescin isothiocyanate conjugated anti-CD4 monoclonal antibody. The sLL preparation is intended to be used as a reference material for CD4+ counting, including the development of higher order reference measurement procedures and has been evaluated in the pilot study CCQM-P102. The sLL reference material was prepared from a pool of three buffy coats from normal human donors, and the PBMCs were separated and labelled with anti-CD4 FITC monoclonal antibody, washed and resuspended in a fixative solution comprised of 10% TransFix.

http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.22614/abstract

2016

Stabilization of Transfected Cells Expressing Low-Incidence Blood Group Antigens: Novel Methods Facilitating Their Use as Reagent-Cells

Sample Type: Cell lines expressing blood group antigens

Analysis Method: Morphological analysis

Authors

C. González, R. Esteban, C. Canals, E. Muñiz-Díaz, N. Nogués

Publication

PLoS ONE (2016) 11(9): e0161968

This paper aimed to generate cell lines expressing blood group antigens (to represent low incidence blood group phenotypes) and to then stabilise these to facilitate their use as reagent cells in clinical laboratories. The addition of TransFix to the cells allowed cell stabilisation and antigen detection for at least 120 days and may facilitate the use of these cells in clinical laboratories, overcoming the limited availability of rare red blood cells. TransFix treated cells were test at 11 time points from Day 0 up to Day 120 showing better morphological preservation (FSC & SSC) vs fresh than lyophilised cells.

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0161968

CSF Stabilisation

2015

Recommendations for quality assurance in multiparametric flow cytometry: first consensus of the Brazilian Group of Flow Cytometry (GBCFLUX)

Analysis Method: Flow Cytometry

Authors

R. P. Correia, A. C. A. Bortolucci, A. C. W. Lopes, A. F. Sandes, A. Paula de Azambuja, M. A. Viana, M. M. Sales, M. Yamamoto, N. S. Bacal

Publication

Jornal Brasileiro de Patologia e Medicina Laboratorial 51(6): 389-396

The Brazilian Group of Flow Cytometry have the objective of contributing to technical and scientific advances in Brazilian clinical and research laboratories. In this publication, they present consensus recommendations to ensure the process quality, technical standardization, and reproducibility of results for all FC working groups in Brazil. For quality control of the pre-analytical phase, they recommend to collect CSF directly into TransFix, which can then be stored at 2-8°C for 48-72 hours.

https://dx.doi.org/10.5935/1676-2444.20150061

The Use of Blood Controls

1999

Absolute CD4+ T-lymphocyte and CD34+ stem cell counts by single platform flow cytometry: the way forward

Authors

D. Barnett, V.Granger, L.Whitby, I.Storie and J.T.Reilly

Publication

British Journal of Haematology (1999) 106: 1059-1062

Stabilised blood controls for CD34 and CD4 were distributed to 280 laboratories worldwide, every 2 months for a year for absolute cell counts. Results were correlated by UK NEQAS. The study showed that the single-platform approach produced consistently lower inter laboratory CV’s for both CD4+ and CD34+ enumeration. Furthermore such technology is likely to prove the method of choice for the quality control of leucodepleted blood products.

http://www.ncbi.nlm.nih.gov/pubmed/10520014

1996

Evaluation of a novel stable whole blood quality control material for lymphocyte subset analysis: Results from the UK NEQAS immune monitoring scheme

Authors

D. Barnett, V.Granger, P.Mayr, I.Storie, G.A.Wilson and J.T.Reilly

Publication

Cytometry (Clinical Cytometry) (1996) 26: 216-222

The suitability of a novel stable whole blood preparation for use as both daily and longitudinal quality control was demonstrated through the UK NEQAS Immune Monitoring scheme. The stabilisation procedure ensures retention of leucocyte light scatter and immunological staining characteristics for up to 300 days. The preparation is also fully compatible with flow cytometer technology, incorporating either whole blood lysis or “no wash, no lyse” techniques. The ranges of inter-laboratory CV’s of the stabilised control were better than those previously obtained with fresh whole blood.

http://www.ncbi.nlm.nih.gov/pubmed/8889394

2001

Low level leucocyte counting: a critical variable in the validation of leucodepleted blood transfusion components as highlighted by an external quality assurance study

Authors

D. Barnett, K. Goodfellow, J. Ginnever, V. Granger, L. Whitby & J.T. Reilly

Publication

Clinical Laboratory Haematology (2001) 23: 43-51

Leucocyte counts of <5 x 106 per blood transfusion product are currently recommended in the UK in order to reduce transfusion related infections and febrile reactions. Routine leucocyte depletion requires the development of reliable internal and external quality assurance programmes. A stabilised low leucocyte blood control manufactured by UK NEQAS was used to determine inter and intra laboratory CV’s. The study highlighted the variability in low level leucocyte counting, especially within the critical range of 5-30 cells/ul. The results highlighted the need for a standardised protocol and nuclear staining reagent for the routine validation of leucocyte depleted blood products.

http://www.ncbi.nlm.nih.gov/pubmed/11422230

1998

Quality assessment of CD34+ stem cell enumeration: experience of the United Kingdom National External Quality Assessment scheme (UK NEQAS) using a unique stable whole blood preparation

Authors

D.Barnett, V.Granger, I.storie, J.Peel, R.Pollitt, T.Smart and J.T.Reilly

Publication

British Journal of Haematology (1998) 102: 553-565

CD34+ peripheral blood stem cell mobilisation and harvesting has replaced autologous bone marrow as a source of stem cells for transplantation. Timing and adequacy of harvests rely upon the accurate enumeration of circulating CD34+ cells. Previous EQA schemes reported inter-laboratory CV’s as high as 284%. A novel stabilised whole blood preparation was distributed to 91 laboratories in 21 countries and participants were required to determine the percentage and absolute values for CD34+ peripheral blood stem cells. Using this method inter-laboratory CV’s were reduced as low as 22% for both percentage and absolute counts.

http://www.ncbi.nlm.nih.gov/pubmed/9695973

2000

Reduction of intra and inter laboratory variation in CD34+ stem cell enumeration using stable test material, standard protocols and targeted training

Authors

D.Barnett, V.Granger, J.Kraan, J.T.Reilly, S.Papa & J.W.Gratama

Publication

British Journal of Haematology (2000) 108: 784-792

The European Working Group on Clinical Cell Analysis has developed a single platform flow cytometric protocol for the enumeration of CD34+ stem cells. A standard protocol was used over 24 clinical sites together using a CD34+ stabilised whole blood control. The use of a common standardisation protocol and targeted training significantly reduced intra and inter laboratory CD34+ cell count variation.

http://www.ncbi.nlm.nih.gov/pubmed/10792284

Standardization of lymphocyte antibody binding capacity – a multi centre study

Authors

D.Barnett, I.Storie, V.Granger, L.Whitbey, J.T.Reilly, S.Brough, S.Garner, J.Lawry, S.Richards, A.E.Bell and B.K.Shenton

Publication

Clinical Laboratory Haematology (2000) 22: 89-96

Quantitative flow cytometry is increasingly being used to characterise non-malignant and malignant disorders, inter and intra laboratory standardisation becomes an important issue. However the lack of standardisation methods and process controls with defined antibody binding values, limits direct comparison. The study showed that a standard protocol is required to achieve low CV’s. Also that stable whole blood can be used as a process control with defined antibody binding capacity values.

http://www.ncbi.nlm.nih.gov/pubmed/10792398


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