Distinguishing cells in a sample by inactivating...

Details Distinguishing cells in a sample by inactivating... Distinguishing cells in a sample by inactivating...

C - Chemistry, Metallurgy

12

Q

C12Q 1/06 (2006.01) C12Q 1/48 (2006.01)

Patent

CA 2607058

A method for detecting the absence or presence of cells of interest in a liquid sample, wherein: (a) the sample: (i) comprises an extracellular medium containing an enzyme with a measurable activity; and (ii) is suspected of containing cells of interest that contain an enzyme with said measurable activity; and (b) the method comprises the steps of: (i) treating the liquid sample with a reagent that inactivates said measurable activity in the extracellular medium, but does not inactivate the measurable activity in said cells of interest; (ii) lysing the cells of interest to release the intracellular enzyme; and (iii) measuring said measurable activity. Thus the intracellular enzyme can be measured without interference from the extracellular enzyme. The invention is particularly useful for treatment of bacterially-infected blood using a detection assay based on adenylate kinase activity.

L'invention concerne un procédé destiné à détecter la présence ou l'absence de cellules d'intérêt dans un échantillon de liquide, dans lequel: (a) l'échantillon: (i) comprend un support extracellulaire contenant une enzyme à activité mesurable; et (ii) est présumé contenir des cellules d'intérêt conenant une enzyme à une activité mesurable,et (b) le procédé comprenant les étapes consistant à: (i) à traiter l'échantillon de liquide avec un réactif qui désactive cette activté mesurable dans le support extracellulaire, mais ne désactive pas l'activité mesurable dans les cellules d'intérêt; (ii) à lyser les cellules d'intérêt afin de libérer l'enzyme intracellulaire; et (iii) à mesurer cette activité mesurable. L'enzyme intracellulaire peut être ainsi mesurée sans interférence de l'enzyme extracellulaire. Cette invention est particulièrement utile dans le traitement de sang infecté par des bactéries à l'aide d'un dosage de détection basé sur une activité d'adénylate kinase.

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