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Reliable and accurate CD4+ T cell count and percent by the portable flow cytometer CyFlow MiniPOC and "CD4 Easy Count Kit-Dry", as revealed by the comparison with the gold standard dual platform technology.

Nasi M, De Biasi S, Bianchini E, Gibellini L, Pinti M, Scacchetti T, Trenti T, Borghi V, Mussini C, Cossarizza A - PLoS ONE (2015)

Bottom Line: The accuracy of CyFlow MiniPOC against Cytomic FC500 showed a correlation coefficient (CC) of 0.98 and 0.97 for CD4+ count and percentage, respectively.Finally, there was no carry-over contamination for samples at all CD4 values, regardless of their position in the sequence of analysis.The cost-effective CyFlow MiniPOC produces rapid, reliable and accurate results that are fully comparable with those from highly expensive dual platform systems.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Medicine, Dentistry and Morphological Sciences, University of Modena and Reggio Emilia School of Medicine, via Campi 287, 41125 Modena, Italy.

ABSTRACT

Background: An accurate and affordable CD4+ T cells count is an essential tool in the fight against HIV/AIDS. Flow cytometry (FCM) is the "gold standard" for counting such cells, but this technique is expensive and requires sophisticated equipment, temperature-sensitive monoclonal antibodies (mAbs) and trained personnel. The lack of access to technical support and quality assurance programs thus limits the use of FCM in resource-constrained countries. We have tested the accuracy, the precision and the carry-over contamination of Partec CyFlow MiniPOC, a portable and economically affordable flow cytometer designed for CD4+ count and percentage, used along with the "CD4% Count Kit-Dry".

Materials and methods: Venous blood from 59 adult HIV+ patients (age: 25-58 years; 43 males and 16 females) was collected and stained with the "MiniPOC CD4% Count Kit-Dry". CD4+ count and percentage were then determined in triplicate by the CyFlow MiniPOC. In parallel, CD4 count was performed using mAbs and a CyFlow Counter, or by a dual platform system (from Beckman Coulter) based upon Cytomic FC500 ("Cytostat tetrachrome kit" for mAbs) and Coulter HmX Hematology Analyzer (for absolute cell count).

Results: The accuracy of CyFlow MiniPOC against Cytomic FC500 showed a correlation coefficient (CC) of 0.98 and 0.97 for CD4+ count and percentage, respectively. The accuracy of CyFlow MiniPOC against CyFlow Counter showed a CC of 0.99 and 0.99 for CD4 T cell count and percentage, respectively. CyFlow MiniPOC showed an excellent repeatability: CD4+ cell count and percentage were analyzed on two instruments, with an intra-assay precision below ± 5% deviation. Finally, there was no carry-over contamination for samples at all CD4 values, regardless of their position in the sequence of analysis.

Conclusion: The cost-effective CyFlow MiniPOC produces rapid, reliable and accurate results that are fully comparable with those from highly expensive dual platform systems.

Show MeSH
Comparison of the CyFlow MiniPOC and CyFlow Counter.Correlation and R2 values for CD4+ T cell counts obtained by Cyflow MiniPOC and CyFlow Counter by using the CD4 easy count kit (A) and the CD4% easy count kit (B).
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pone.0116848.g001: Comparison of the CyFlow MiniPOC and CyFlow Counter.Correlation and R2 values for CD4+ T cell counts obtained by Cyflow MiniPOC and CyFlow Counter by using the CD4 easy count kit (A) and the CD4% easy count kit (B).

Mentions: The values of CD4+ T cell count obtained by CyFlow MiniPOC showed a very good correlation with those obtained with the CyFlow Counter either by using the “CD4 Easy Count Kit” (Fig. 1A; R2 = 0.963, correlation coefficient 0.98) or by using the “CD4% Easy Count Kit” (Fig. 1B; R2 = 0.979, correlation coefficient 0.99). The Bland-Altman plot indicates that the overall, absolute CD4+ T cell counts obtained by the two methods were in excellent agreement (Fig. 2A and 2B). The values of CD4+ T cell percent obtained by CyFlow MiniPOC showed a strong correlation with those obtained with the CyFlow Counter by using the CD4% easy count kit (Fig. 3A; R2 = 0.976, correlation coefficient 0.99). All individual data points represented in Figs. 1–3 are shown in S1 Table.


Reliable and accurate CD4+ T cell count and percent by the portable flow cytometer CyFlow MiniPOC and "CD4 Easy Count Kit-Dry", as revealed by the comparison with the gold standard dual platform technology.

Nasi M, De Biasi S, Bianchini E, Gibellini L, Pinti M, Scacchetti T, Trenti T, Borghi V, Mussini C, Cossarizza A - PLoS ONE (2015)

Comparison of the CyFlow MiniPOC and CyFlow Counter.Correlation and R2 values for CD4+ T cell counts obtained by Cyflow MiniPOC and CyFlow Counter by using the CD4 easy count kit (A) and the CD4% easy count kit (B).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4306486&req=5

pone.0116848.g001: Comparison of the CyFlow MiniPOC and CyFlow Counter.Correlation and R2 values for CD4+ T cell counts obtained by Cyflow MiniPOC and CyFlow Counter by using the CD4 easy count kit (A) and the CD4% easy count kit (B).
Mentions: The values of CD4+ T cell count obtained by CyFlow MiniPOC showed a very good correlation with those obtained with the CyFlow Counter either by using the “CD4 Easy Count Kit” (Fig. 1A; R2 = 0.963, correlation coefficient 0.98) or by using the “CD4% Easy Count Kit” (Fig. 1B; R2 = 0.979, correlation coefficient 0.99). The Bland-Altman plot indicates that the overall, absolute CD4+ T cell counts obtained by the two methods were in excellent agreement (Fig. 2A and 2B). The values of CD4+ T cell percent obtained by CyFlow MiniPOC showed a strong correlation with those obtained with the CyFlow Counter by using the CD4% easy count kit (Fig. 3A; R2 = 0.976, correlation coefficient 0.99). All individual data points represented in Figs. 1–3 are shown in S1 Table.

Bottom Line: The accuracy of CyFlow MiniPOC against Cytomic FC500 showed a correlation coefficient (CC) of 0.98 and 0.97 for CD4+ count and percentage, respectively.Finally, there was no carry-over contamination for samples at all CD4 values, regardless of their position in the sequence of analysis.The cost-effective CyFlow MiniPOC produces rapid, reliable and accurate results that are fully comparable with those from highly expensive dual platform systems.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Medicine, Dentistry and Morphological Sciences, University of Modena and Reggio Emilia School of Medicine, via Campi 287, 41125 Modena, Italy.

ABSTRACT

Background: An accurate and affordable CD4+ T cells count is an essential tool in the fight against HIV/AIDS. Flow cytometry (FCM) is the "gold standard" for counting such cells, but this technique is expensive and requires sophisticated equipment, temperature-sensitive monoclonal antibodies (mAbs) and trained personnel. The lack of access to technical support and quality assurance programs thus limits the use of FCM in resource-constrained countries. We have tested the accuracy, the precision and the carry-over contamination of Partec CyFlow MiniPOC, a portable and economically affordable flow cytometer designed for CD4+ count and percentage, used along with the "CD4% Count Kit-Dry".

Materials and methods: Venous blood from 59 adult HIV+ patients (age: 25-58 years; 43 males and 16 females) was collected and stained with the "MiniPOC CD4% Count Kit-Dry". CD4+ count and percentage were then determined in triplicate by the CyFlow MiniPOC. In parallel, CD4 count was performed using mAbs and a CyFlow Counter, or by a dual platform system (from Beckman Coulter) based upon Cytomic FC500 ("Cytostat tetrachrome kit" for mAbs) and Coulter HmX Hematology Analyzer (for absolute cell count).

Results: The accuracy of CyFlow MiniPOC against Cytomic FC500 showed a correlation coefficient (CC) of 0.98 and 0.97 for CD4+ count and percentage, respectively. The accuracy of CyFlow MiniPOC against CyFlow Counter showed a CC of 0.99 and 0.99 for CD4 T cell count and percentage, respectively. CyFlow MiniPOC showed an excellent repeatability: CD4+ cell count and percentage were analyzed on two instruments, with an intra-assay precision below ± 5% deviation. Finally, there was no carry-over contamination for samples at all CD4 values, regardless of their position in the sequence of analysis.

Conclusion: The cost-effective CyFlow MiniPOC produces rapid, reliable and accurate results that are fully comparable with those from highly expensive dual platform systems.

Show MeSH