Comparative assessment of five alternative methods for CD4+ T-lymphocyte enumeration for implementation in developing countries.

This study compares five alternative methods with regard to the accuracy of CD4+ cell counts measured at the same time by standard flow cytometry (FC). The alternative methodologies studied included: FACSCount; VCS Technology/Coulter Cytospheres; Dynabeads MPC-Q; Capcellia; and Opti-CIM. These methods were performed simultaneously on the blood samples of 40 HIV-infected patients and 20 healthy adult employees collected on 4 separate days. Overall findings revealed that the correlation coefficients between results obtained by FC and each alternative method were similar whatever the range of CD4+ cell count (> or < 500/cu. mm) except for Capcellia. The median differences between absolute numbers of CD4+ T lymphocytes obtained by standard FC and each alternative method were 45 (n = 45) for FACSCount 56 (n = 46) for Dynabeads 195 (n = 55) for Cytospheres 134 (n = 49) for Capcellia and 126 (n = 27) for Opti-CIM. The study showed that FACSCount equipment was lowest compared with the other methods with regard to the median difference in absolute numbers of CD4+ T lymphocytes. Moreover it was also determined that with Dynabeads when compared with the standard FC method an excellent correlation coefficient was found without regard to the CD4+ cell number. In conclusion the FACSCount and Dynabeads methods compared well with standard FC irrespective of CD4+ cell numbers. The Dynabeads method appears to be less expensive hence it has potential in countries with limited economic resources.

[1]  L. Brammer,et al.  Influenza surveillance--United States, 1992-93 and 1993-94. , 1997, MMWR. CDC surveillance summaries : Morbidity and mortality weekly report. CDC surveillance summaries.

[2]  J V Giorgi,et al.  Application of flow cytometry to the study of HIV infection. , 1990, AIDS.

[3]  B. Pau,et al.  Whole blood capcellia CD4/CD8 immunoassay for enumeration of CD4+ and CD8+ peripheral T lymphocytes. , 1999, Clinical chemistry.

[4]  I. Lisse,et al.  Determination of CD4+ and CD8+ lymphocytes with the cytosphere assay: a comparative study with flow cytometry and the immunoalkaline phosphatase method. , 1995, Clinical immunology and immunopathology.

[5]  P. Paoli,et al.  Two-site enzyme immunoassay of CD4 and CD8 molecules on the surface of T lymphocytes from healthy subjects and HIV-1-infected patients. , 1994, Clinical chemistry.

[6]  T. Denny,et al.  Evaluation of a method for counting absolute numbers of cells with a flow cytometer , 1997, Clinical and diagnostic laboratory immunology.

[7]  K. Pallangyo,et al.  Evaluation of the FACScount, TRAx CD4 and Dynabeads methods for CD4 lymphocyte determination. , 1996, Journal of immunological methods.

[8]  H. Dahourou,et al.  Evaluation of a quantitative determination of CD4 and CD8 molecules as an alternative to CD4 + and CD8 + T lymphocyte counts in Africans , 1999, Tropical medicine & international health : TM & IH.

[9]  T. Quinn,et al.  A manual bead assay for the determination of absolute CD4+ and CD8+ lymphocyte counts in human immunodeficiency virus-infected individuals , 1995, Clinical and diagnostic laboratory immunology.