Evaluation of a dry format reagent alternative for CD4 T-cell enumeration for the FACSCount system: a report on a Moroccan-Canadian study.
BACKGROUND
Efforts to improve alternative CD4 T-cell counting methods are critical to accelerate the implementation of HIV antiretroviral therapy in resources limited regions. Substituting liquid format reagents to eliminate cold-chain transportation and refrigerated storage with dry format reagents contributes to higher efficiency supply management solution especially for laboratories at remote locations. ReaMetrix has developed dry format reagent kits compatible with the FACSCount system, a dedicated flow cytometer for T-cell subset enumeration widely used in resource limited settings. A dual site collaborative study was designed to compare T-cell subsets using both the new dry format ReaMetrix reagent and the original BD Biosciences liquid reagents.


METHODS
A total of 167 HIV positive samples prepared with Rea T Count (ReaMetrix) and FACSCount (BD Biosciences) reagents were analyzed using FACSCount Systems. To compare both methods, Bland-Altman, Pollock, Scott % similarity and correlation coefficient statistical analysis was applied. Immuno-Trol served as an assay processing control and quality indicator of interlaboratory and intralaboratory variation.


RESULTS
The mean bias and limits of agreement for CD4 T-cell measurements between Rea T Count and FACSCount reagents were -16 cells/microl (-4.6%) and -74 to +43, respectively. The correlation obtained was 0.988 with a similarity of 97.9%. Between laboratory variation data was very good with %CV below 10%.


CONCLUSIONS
The introduction of dry reagents permits the elimination of cold-chain transportation and the on-site refrigerated storage without compromise to assay quality. The substitution of dry reagents facilitates easier supply management practice that will assure wider access to quality HIV treatment.
National HIV Immunology Laboratory, Public Health Agency of Canada, Ottawa, Ontario, Canada K1A 0K9. michele_bergeron@phac-aspc.gc.ca
Michèle
M
Bergeron
Tao
T
Ding
Elmir
E
Elharti
Hicham
H
Oumzil
Nadia
N
Soucy
Hicham
H
Harmouche
Saad
S
Chaouch
Rajae El
RE
Aouad
Christian
C
Chabot
Francis
F
Mandy
D015703:Antigens, CD / Q000276:immunology; D018791:CD4 Lymphocyte Count / Q000379:methods*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000473:pathology / Q000821:virology; D002170:Canada; D005434:Flow Cytometry / Q000379:methods*; D015658:HIV Infections / Q000473:pathology / Q000821:virology; D006801:Humans; D009018:Morocco; D000091402:Prohibitins; D011786:Quality Control; D011933:Reagent Kits, Diagnostic
D015703:Antigens, CD; C000717331:PHB2 protein, human; D000091402:Prohibitins; D011933:Reagent Kits, Diagnostic