Investigating the use of standardized EuroFlow flow cytometry panels for thecharacterisation and diagnosis of Chronic lymphocyticleukaemia in the Tygerberg Academic Hospital, South Africa.

[摘要] ENGLISH ABSTRACT : Background: Flow cytometry (FC) immunophenotyping is crucial in the diagnosis andclassification standardisation of haematological malignancies. FC techniques requirestandardisation to produce reliable and reproducible results which are important ininter-laboratory studies for laboratory methodology improvement. The aim of this studyis to introduce standardised multicolour FC in the diagnosis of haematologicalmalignancies using chronic lymphocytic leukaemia (CLL) as the proof of principle. Inaddition, we aim to document the incidence of CLL from the year 2011 to 2016 in theTygerberg Academic Hospital (TAH) catchment area of Cape Town, South Africa (SA).Methods: Twenty CLL patients were recruited at TAH. Bio-specimens were preparedand analysed on the Beckman Coulter Navios flow cytometer using Euroflow™standardised FC protocols and immunophenotypic panels with two tubes for detectingB-cell chronic lymphoproliferative disorders (B-CLPD). Tube 1 included CD20, CD4,CD45, CD8, lg-Kappa, CD56, lg-Lambda, CD5, CD19, TCRyσ, CD3 and CD38. Tube2 included CD20, CD45, CD23, CD10, CD79b, CD19, CD200 and CD43. Combined,the two tubes identified CLL from other B-CLPD. The CLL immunophenotypic profileswere stored in a database using the compass tool of the Infinicyt™ FC software. Inaddition, the clinical records of patients diagnosed with CLL at TAH over a 6-yearperiod from the year 2011 to 2016 were retrieved and analysed using descriptivestatistics.Results: In comparison with the SA National Health Laboratory Service (NHLS) resultsat TAH, the Euroflow™ standardised multicolour FC panels and protocols are suitablefor immunophenotyping CLL in this SA population. An immunophenotype database for20 CLL diagnosed at TAH was constructed using the EuroFlow™ standardisedmulticolour FC panels. For the epidemiology part of the study, a total of 80 CLL patientswere studied. There were slightly more females (51.2%) and the mean age at diagnosiswas 67 years (37 to 95). Ninety one percent of the patients were aged 50 years andabove. Males presented with the disease at a younger age (mean 63 years) thanfemales (mean 70 years). CLL concurrent with HIV was not common (4%) and thesepatients were younger than 50 years. Twenty-one patients were tested forchromosomal aberrations trisomy 12 and deletion 11q, 24% and 33% were positiverespectively. Deletion 13q was assessed in 25 patients and 16% were positive. Twenty patients were tested for deletion 17p and all were negative. Translocations t(8;14),t(11;14) and t (14;18) were negative in 1, 8 and 4 patients respective.Discussion: Accurate and consistent laboratory techniques and strict standardisationin FC enhances the confidence in inter-laboratory studies. Establishment ofhaematological malignancy immunophenotype databases would allow for fasterdifferential diagnoses of new disease cases which is needed within our setting.Furthermore, these databases permit clear identification of atypical cases. Monitoringhaematological malignancy trends is a crucial step in the management of the disease.

[发布机构] Stellenbosch University