Barcelona 2013
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| Abstract #30 - E-Posters English | ||||
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Session: 50.10: E-Posters English (Poster) on Sunday in Chaired by Authors: Presenting Author: Mr Sumit Gupta - Kasturba Medical College, India |
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| Additional Authors: Dr. Jordi Casabona, Sra Cristina Sanclemente, Dra. Anna Esteve, Dra. Victoria Gonzalez, Grupo HIVITS TS, | ||||
| Aim: To study and compare studied dual-platform protocols utilizing only 2 essential antibodies for the CD4 cell enumeration. | ||||
| Method / Issue: CD4+ cell counts to monitor HIV/AIDS patients are essential in the era of highly active antiretroviral therapy. Complex flow cytometric (FCM) assays being expensive, cost-reduction in these assays is an attractive goal for under-resourced laboratories. We retro-prospectively studied 2 previously described dual-platform protocols utilizing only 2 essential antibodies for their applicability to an Indian setting. Archival list mode data files, hematological indices and final reports of 673 samples tested over 2003-2010 using a standardized method (CYTO-STAT tetraCHROME CD45-FITC / CD4-RD1 / CD8-ECD / CD3-PC5) on an EPICS-XL machine (Beckman Coulter, FL, USA) were retrieved. These were re-analyzed using 2 informative antibodies (CD45 and CD4) on KaluzaTM software (Beckman Coulter). Sequential gating calculated CD4+ T-cells as percentage subsets of all lymphocytes (low side scatter and bright CD45 expression) as well as of all leukocytes (panleukogating). Absolute lymphocyte and total leukocyte counts were obtained on LH750 analyzers (Beckman Coulter). CD4 counts reported by the 4-color (historical) method were compared with the lymphocyte-gated and panleukogated results using SPSS v17.0 statistical software | ||||
| Results / Comments: Absolute CD4+ T cell counts ranged from 0 to 4196 cells/μL. Bland Altman statistics revealed no significant bias (10.04 cells/μL, 95% agreement limits -95.3 to 115.31) for lymphocyte-gated protocol and some underestimation (bias=-87.77cells/μL, 95% agreement limits -698.78 to 523.38) by the panleukogating protocol. Paired sample tests showed mean difference of 10.04 cells/μL between the conventional and the lymphocyte gated counts. At a 200 cells/μL cutoff, the sensitivity and specificity of lymphocyte gating method were 100% & 99.6%, and that of panleukogating were 88.1% and 99.8% respectively. | ||||
| Discussion: We found that enumerating CD4+ T cells by omitting non-essential antibodies (using CD45/CD4 instead of CD45/CD3/CD4/CD8) by a dual-platform technique is feasible and accurate. Lymphocyte-gating is superior to the panleukogating method in this regard. | ||||
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