Comparison of three methods of CD34(+) cell enumeration in peripheral blood: dual-platform ISHAGE protocol versus single-platform, versus microvolumefluorimetry
P. Chapple et al., Comparison of three methods of CD34(+) cell enumeration in peripheral blood: dual-platform ISHAGE protocol versus single-platform, versus microvolumefluorimetry, CYTOTHERAPY, 2(5), 2000, pp. 371-376
Citations number
16
Categorie Soggetti
Hematology,"Medical Research Diagnosis & Treatment
Background Quantitation of peripheral blood (PB) CD34(+) cells is now nn es
tablished method for timing PBPC harvesting. Recent refinements to the dual
-platform ISHAGE gating strategy for CD34(+) cells has seen the introductio
n of microbeads to enable absolute counting of cells on a single instrument
platform. This eliminates the need for total WBCC performed on an automate
d hematology analyzer and potentially increases the analytical precision of
the methodology! At the same time, alternative methods for CD34(+) cell en
umeration have started to emerge, notably microvolume fluorimetry, which fo
rms the basis of the fully-automated STELLer CD34 method using the Imagn 20
00.
Methods We performed a three-way evaluation of these methods. Sixty-eight s
amples of PB from 42 patients undergoing PBPC mobilization weve analyzed by
all three methods and correlations between all three calculated. The two-p
latform ISHAGE method was used as the reference method.
Results Precision and linearity of the single-platform and STELLer CD34 ass
ays were excellent Correlation with the dual-platform reference method was
also excellent (single-platform method slope = 1.03, intercept = - 0.03 and
R-2 = 0.9325 STELLer CD34 assay slope = 0.827 intercept = 4.27 R-2 = 0.821
5). Bias, determined by Bland-Altman analysis, was 1.16 and -1.62 for singl
e platform and STELLer CD34 assay respectively.
Conclusion The three methods of CD34(+) cell enumeration gave equivalent re
sults. The single-platform methodology negated the need for a separate whit
e cell analyzer while the STELLer CD34 methodology was technically the simp
lest.