Method for the enumeration of micronucleated erythrocyte...

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G01N 33/80 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2529802

A method for the enumeration of micronucleated erythrocyte populations while distinguishing platelet and platelet-associated aggregates involves the use of a first fluorescent labeled antibody having binding specificity for a surface marker for reticulocytes, a second fluorescent labeled antibody having binding specificity for a surface marker for platelets, and a nucleic acid staining dye that stains DNA (micronuclei) in erythrocyte populations. Because the fluorescent emission spectra of the first and second fluorescent labeled antibodies do not substantially overlap with one another or with the emission spectra of the nucleic acid staining dye, upon excitation of the labels and dye it is possible to detect the fluorescent emission and light scatter produced by the erythrocyte populations and platelets, and count the number of cells from one or more erythrocyte populations in said sample. In particular, the use of the second antibody prevents interference by platelet-associated aggregates in the scoring procedures.

Procédé de numération de populations d'érythrocytes à micronoyaux avec distinction des plaquettes et des amas associés aux plaquettes, qui consiste à utiliser un premier anticorps marqué fluorescent ayant une spécificité de liaison pour un marqueur de surface des réticulocytes, un second anticorps marqué fluorescent ayant une spécificité de liaison pour un marqueur de surface des plaquettes, et un colorant d'acide nucléique qui colore l'ADN (micronoyaux) dans les populations d'érythrocytes. Du fait que les spectres d'émission fluorescents des premier et second anticorps fluorescents marqués ne se chevauchent pas complètement l'un l'autre ou ne se chevauchent pas complètement avec le spectre d'émission du colorant d'acide nucléique, il est possible, après excitation des marqueurs et du colorant, de détecter l'émission fluorescente et la diffusion de la lumière produites par les populations d'érythrocytes et de plaquettes et de compter le nombre de cellules d'une ou plusieurs populations d'érythrocytes dans un échantillon. En particulier, l'utilisation du second anticorps empêche l'interférence provoquée par des amas associés aux plaquettes dans les procédures de comptage.

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