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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.

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Related in: MedlinePlus

Carry-over contamination analysis.The CD4 absolute count (A) and CD4% (B) were plotted as the mean value of three analysis sequences. Error bars indicate the standard deviation of three measurements.
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pone.0116848.g007: Carry-over contamination analysis.The CD4 absolute count (A) and CD4% (B) were plotted as the mean value of three analysis sequences. Error bars indicate the standard deviation of three measurements.

Mentions: As shown in Fig. 7, using the CyFlow MiniPOC, we observed no significant differences among all samples with low CD4 count and percentage, all those with medium CD4 count and percentage and all those with high CD4 count and percentage, regardless of their position in the sequence of analysis. All individual data points represented in Fig. 7 are shown in S4 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)

Carry-over contamination analysis.The CD4 absolute count (A) and CD4% (B) were plotted as the mean value of three analysis sequences. Error bars indicate the standard deviation of three measurements.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0116848.g007: Carry-over contamination analysis.The CD4 absolute count (A) and CD4% (B) were plotted as the mean value of three analysis sequences. Error bars indicate the standard deviation of three measurements.
Mentions: As shown in Fig. 7, using the CyFlow MiniPOC, we observed no significant differences among all samples with low CD4 count and percentage, all those with medium CD4 count and percentage and all those with high CD4 count and percentage, regardless of their position in the sequence of analysis. All individual data points represented in Fig. 7 are shown in S4 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
Related in: MedlinePlus