Accuracy of domestic T lymphocytes reagent and flow cytometry: an applicative evaluation

  Objective  To evaluate the accuracy of a domestic reagent (Tongsheng reagent) for determination of absolute count of cluster of differentiation 4 positive (CD4⁺) T lymphocytes with different multicolor flow cytometry.

  Methods  Parallel tests for accuracy in absolute counts of T lymphocyte subgroups detected using domestic reagents and instrument were performed on whole blood samples of 206 human immunodeficiency virus (HIV) infected people during July 2014. The domestic reagents used in the detection were Tongsheng three color (CD3-FITC/CD8-PE/CD4-APC CD3/8/4/CD3-FITC/CD4-PE /CD45-PerCP CD3/4/45) and Tongsheng four color (CD3-FITC/CD8-PE/CD45-PerCP/CD4-APC CD3/8/45/4) and the domestic instrument was ACEA NovoCyte flow cytometry. The reference reagent and instrument were BD four color reagent kit and ral BD FACSCalibur flow cytometry.

  Results  For all the blood samples, the correlation coefficients between the results from detections with domestic reagents and instrument and reference reagent and instrument were all greater than 0.96. Compared with the results using reference reagent and instrument, when using Tongsheng three color (CD3/8/4 or CD3/4/45) and Tongsheng four color (CD3/8/45/4) reagents and ACEA NovoCyte flow cytometry, the mean deviations of absolute counts (cell/μL) were 2.49 ± 50.5, – 3.1 ± 43.8 and – 6.2 ± 58.2 for CD4; 10.3 ± 115.3, – 1.7 ± 108.0 and 14.3 ± 125.4 for CD3; – 4.6 ± 78.6 and – 19.6 ± 87.2 for CD8, respectively. No significant differences were observed between the absolute counts of CD4, CD3, and CD8 T lymphocytes determined with domestic and reference reagents and instrument.

  Conclusion  Domestic Tongsheng three color and four color reagents and ACEA NovoCyte can be used for accurate determinations of T lymphocytes absolute counts.