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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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Partec CyFlow MiniPOC.The Partec CyFlow MiniPOC counting device (A) and its display (B) in routine (left) or expert mode (right). Both images are used with permission of Partec/Sysmex GmbH, Germany.
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pone.0116848.g008: Partec CyFlow MiniPOC.The Partec CyFlow MiniPOC counting device (A) and its display (B) in routine (left) or expert mode (right). Both images are used with permission of Partec/Sysmex GmbH, Germany.

Mentions: The main aim of our study was to compare a system that fulfills the ASSURED criteria, such as that formed by the CyFlow MiniPOC and the CD4% Count Dry-Kit (shown in Fig. 8A), with the gold standard, dual platform system that is routinely used in an Italian Hospital since several years, and with another single-platform low cost system, i.e. the CyFlow Counter. We found that the results obtained with the CyFlow miniPOC with the use of the CD4% count dry kit are comparable either to those obtained with an independent reference system, i.e. the Beckman Coulter FC500 plus Coulter HmX, or to the single platform CyFlow Counter. Furthermore, the results obtained with the dry kits designed for the CyFlow miniPOC were fully comparable with the liquid kits “CD4 easy count kit” and “CD4% easy count kit” used with the CyFlow Counter.


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)

Partec CyFlow MiniPOC.The Partec CyFlow MiniPOC counting device (A) and its display (B) in routine (left) or expert mode (right). Both images are used with permission of Partec/Sysmex GmbH, Germany.
© Copyright Policy
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC4306486&req=5

pone.0116848.g008: Partec CyFlow MiniPOC.The Partec CyFlow MiniPOC counting device (A) and its display (B) in routine (left) or expert mode (right). Both images are used with permission of Partec/Sysmex GmbH, Germany.
Mentions: The main aim of our study was to compare a system that fulfills the ASSURED criteria, such as that formed by the CyFlow MiniPOC and the CD4% Count Dry-Kit (shown in Fig. 8A), with the gold standard, dual platform system that is routinely used in an Italian Hospital since several years, and with another single-platform low cost system, i.e. the CyFlow Counter. We found that the results obtained with the CyFlow miniPOC with the use of the CD4% count dry kit are comparable either to those obtained with an independent reference system, i.e. the Beckman Coulter FC500 plus Coulter HmX, or to the single platform CyFlow Counter. Furthermore, the results obtained with the dry kits designed for the CyFlow miniPOC were fully comparable with the liquid kits “CD4 easy count kit” and “CD4% easy count kit” used with the CyFlow Counter.

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