Effects of Storage: Whole Blood Specimens for CD4-T Lymphocytes Determination in Yaoundé, Cameroon

Marie Claire Okomo Assoumou, George Nguefack, Graeme Brendon Jacobs, Elise Guidem, Gilbert Ndeh Doh, Martha Tongo Mesembe, Emilia Lyonga, George Mondinde Ikomey

Abstract


ABSTRACT
Introduction. Stored blood specimens collected in Ethylene Diamine Tetra Acetic Acid (EDTA) anti-coagulant tubes for CD4 T-Cell determinations are inevitable practices in Cameroon and many other countries worldwide. This study aimed to evaluate the reproducibility of results obtained from blood samples stored at room temperature (25° C) and 4° C within six days post collection. Methods. Whole blood samples collected in EDTA collection tubes were analyzed for CD4 absolute counts using the FACS Count Machine (Becton Dickinson, France). We Used the FACS Count reagent kit (CD4, CD8 and CD3 combine). Samples were strictly analyzed using the procedural manual of the kit .Freshly collected blood samples in day one served as control. Results. There was a significant correlation with results obtained at day 2 and 3 (47 and 72 hours).Day 4, 5, 6 (72, 96 and 120 hours) did not show any correlation with day one. Blood samples stored at -4°C degrees did not correlate with any other day. Conclusion. Reproducibility of CD4 cell count could only be evaluated up to 72 hours, whereas blood samples stored in the fridge at -4°C were not reproducible after day one. Our findings can serve as a guide to clinical laboratories and health care providers handling samples in rural and urban settings.

RÉSUMÉ
Introduction. La conservation des échantillons de sang prélevés dans les tubes contenant l’anticoagulant Ethylène Diamine-Tétra-Acétique (EDTA) pour la détermination du taux de lymphocytes T CD4 sont des pratiques courantes au Cameroun et dans de nombreux pays du monde .Cette étude visait à évaluer la reproductibilité des résultats obtenus à partir des échantillons de sang stockés à température ambiante (25°C) dans les six jours suivant la collecte. Méthodologie. Les échantillons de sang total collectés dans des tubes EDTA ont été analysés afin de déterminer le nombre absolu de CD4 en utilisant l’appareil FACS Count (Becton Dickinson, France).Nous avons utilisé le kit de réactifs FACS Count (CD4, CD8 et CD3 combinés). Les échantillons ont été analysés en respectant la procédure décrite par le fabriquant du kit. Les échantillons de sang fraîchement prélevés au jour 1 ont servi de témoins. Résultats. Nous avons trouvé une corrélation significative entre les résultats obtenus au jour 2 et 3 (47 et 72 heures). Il n’y avait pas de corrélation entre les jours 4, 5, 6 (72, 96 et 120 heures) et le jour un. Les résultats des échantillons de sang stockés à - 4°C n’ont été corrélés avec ceux d’aucun autre jour. Conclusion. La reproductibilité du nombre de cellules CD4 ne peut être évaluée que jusqu'à 72 heures, alors que les échantillons de sang stockés dans le réfrigérateur à - 4°C ne sont pas reproductibles après le premier jour. Nos résultats peuvent servir de guide aux laboratoires cliniques et aux prestataires de soins de santé manipulant des échantillons en milieu rural et urbain.


Keywords


CD4-T Lymphocytes, Whole blood, Cameroon

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