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Panleucogating as an accurate and affordable flow cytometric protocol to analyse lymphocyte subsets among HIV-positive patients on HAART treatment in Mozambique.
J Biol Regul Homeost Agents. 2005 Jul-Dec; 19(3-4):169-75.JB

Abstract

In Africa tens of millions of people are HIV+. Prevention alone is not effective, and needs to be coupled with anti-retroviral treatment (HAART). Laboratory tests as CD4+ T cell count are fundamental tools in HIV disease monitoring, but they require costly equipment, reagents and specialised manpower. The goal of this study was to minimise and optimise the reagents needed for a reliable routine CD4+ cell count in a resource-poor setting (Mozambique). Panleucogating protocol (PLG), requires two antibodies only, CD45 and CD4, or three if CD8 is requested for special clinical reasons. PLG was compared with the current protocol used in two Mozambique hospitals, based on FSC/SSC gating and CD3/CD4/CD8 staining. 189 samples from HIV+ patients, included in the Community of Sant'Egidio's DREAM program and on HAART were processed with both protocols. The overall correlation of the lymphocyte subsets measurements was satisfactory, with r2 always >0.96. The Bland-Altman analysis of CD4+ cell count showed a negative bias when CD4+ cells were <15%, due to the imprecise FSC/SSC gating used previously. When CD4+ cells were >15% the negative bias tended to zero, further confirming the better quality of the PLG gating strategy. Two- or three color PLG protocol, in double platform, currently seems the most accurate and affordable method to monitor CD4+ lymphocytes and CD4/CD8 ratio by flow cytometry in resource-poor medical settings.

Authors+Show Affiliations

Department of Oncology, Division of Surgical, Molecular and Ultrastructural Pathology, University of Pisa and Pisa University Hospital, Italy. susannaceffa@hotmail.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16602633

Citation

Ceffa, S, et al. "Panleucogating as an Accurate and Affordable Flow Cytometric Protocol to Analyse Lymphocyte Subsets Among HIV-positive Patients On HAART Treatment in Mozambique." Journal of Biological Regulators and Homeostatic Agents, vol. 19, no. 3-4, 2005, pp. 169-75.
Ceffa S, Erba F, Assane M, et al. Panleucogating as an accurate and affordable flow cytometric protocol to analyse lymphocyte subsets among HIV-positive patients on HAART treatment in Mozambique. J Biol Regul Homeost Agents. 2005;19(3-4):169-75.
Ceffa, S., Erba, F., Assane, M., Coelho, E., Calgaro, M., & Brando, B. (2005). Panleucogating as an accurate and affordable flow cytometric protocol to analyse lymphocyte subsets among HIV-positive patients on HAART treatment in Mozambique. Journal of Biological Regulators and Homeostatic Agents, 19(3-4), 169-75.
Ceffa S, et al. Panleucogating as an Accurate and Affordable Flow Cytometric Protocol to Analyse Lymphocyte Subsets Among HIV-positive Patients On HAART Treatment in Mozambique. J Biol Regul Homeost Agents. 2005 Jul-Dec;19(3-4):169-75. PubMed PMID: 16602633.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Panleucogating as an accurate and affordable flow cytometric protocol to analyse lymphocyte subsets among HIV-positive patients on HAART treatment in Mozambique. AU - Ceffa,S, AU - Erba,F, AU - Assane,M, AU - Coelho,E, AU - Calgaro,M, AU - Brando,B, PY - 2006/4/11/pubmed PY - 2006/6/6/medline PY - 2006/4/11/entrez SP - 169 EP - 75 JF - Journal of biological regulators and homeostatic agents JO - J Biol Regul Homeost Agents VL - 19 IS - 3-4 N2 - In Africa tens of millions of people are HIV+. Prevention alone is not effective, and needs to be coupled with anti-retroviral treatment (HAART). Laboratory tests as CD4+ T cell count are fundamental tools in HIV disease monitoring, but they require costly equipment, reagents and specialised manpower. The goal of this study was to minimise and optimise the reagents needed for a reliable routine CD4+ cell count in a resource-poor setting (Mozambique). Panleucogating protocol (PLG), requires two antibodies only, CD45 and CD4, or three if CD8 is requested for special clinical reasons. PLG was compared with the current protocol used in two Mozambique hospitals, based on FSC/SSC gating and CD3/CD4/CD8 staining. 189 samples from HIV+ patients, included in the Community of Sant'Egidio's DREAM program and on HAART were processed with both protocols. The overall correlation of the lymphocyte subsets measurements was satisfactory, with r2 always >0.96. The Bland-Altman analysis of CD4+ cell count showed a negative bias when CD4+ cells were <15%, due to the imprecise FSC/SSC gating used previously. When CD4+ cells were >15% the negative bias tended to zero, further confirming the better quality of the PLG gating strategy. Two- or three color PLG protocol, in double platform, currently seems the most accurate and affordable method to monitor CD4+ lymphocytes and CD4/CD8 ratio by flow cytometry in resource-poor medical settings. SN - 0393-974X UR - https://www.unboundmedicine.com/medline/citation/16602633/Panleucogating_as_an_accurate_and_affordable_flow_cytometric_protocol_to_analyse_lymphocyte_subsets_among_HIV_positive_patients_on_HAART_treatment_in_Mozambique_ L2 - http://www.diseaseinfosearch.org/result/9735 DB - PRIME DP - Unbound Medicine ER -