Methods and devices for detecting MxA in a sample on a sample anaysis device
Devices and methods incorporate mucolytic agents into a point-of-care testing device. The sample is loaded, and then the sample travels until it encounters one or more lysis agents and/or mucolytic agents. The mucolytic agent is preferably pre-loaded onto the collection device. In a preferred embodiment, the mucolytic agent is localized between the sample application zone and the conjugate zone. In embodiments with a sample compressor, one or more mucolytic agents may be pre-loaded and dried on the sample compressor, the sample collector, in various locations on the test strip, or in the running buffer.
1 . A method for detecting MxA in a sample on a sample analysis device that includes a flow of an elution medium from an elution medium application zone to a detection zone, comprising:
a) transferring a blood sample that includes lymphocyte cells onto a sample application zone on the sample analysis device;
b) applying an elution medium that includes a lysis agent capable of lysing lymphocyte cells to the elution medium application zone, such that the elution medium, including the at least one lysis agent, flows through the sample application zone toward the detection zone to produce lysed lymphocyte cells, thereby releasing intracellular MxA present within the lymphocyte cells into the elution medium prior to reaching the detection zone;
c) reducing components of the lysed lymphocytes at a blocking zone of the sample analysis device that are larger than a pore size of a barrier at the blocking zone, wherein the blocking zone is positioned between the sample application zone and the detection zone; and
d) detecting the presence of MxA at the detection zone on the sample analysis device.
2 . The method of claim 1 , wherein a conjugate zone is located between the sample application zone and the detection zone.
3 . The method of claim 2 , wherein the blocking zone comprises pores that act as a barrier to slow or arrest lysed materials effectively larger than the pore size of the pores, wherein the blocking zone is positioned between the sample application zone and the conjugate zone.
4 . The method of claim 2 , wherein the blocking zone is positioned between the sample application zone and the conjugate zone.
5 . The method of claim 1 , wherein the sample is whole blood.
6 . The method of claim 1 , further comprising the step of detecting a presence of an extracellular protein at the detection zone.
7 . The method of claim 1 , wherein the sample analysis device is a lateral flow immunoassay device.
8 . The method of claim 1 , wherein the sample analysis device comprises a chromatography test strip.
9 . The method of claim 1 , wherein the blocking zone comprises pores that act as a barrier to slow or arrest lysed materials from the lysed lymphocyte cells effectively larger than the pore size of the barrier.
10 . The method of claim 1 , wherein the barrier is a physical barrier.
11 . The method of claim 1 , wherein erythrocytes of the blood sample are physically removed by the barrier to enhance selective lysis of leukocytes of the blood sample.
12 . The method of claim 1 , further comprising:
(i) exposing the MxA to a chemiluminescent detection reagent to form a detectable binding complex comprising the MxA coupled to the chemiluminescent detection reagent; and
(ii) detecting a chemiluminescence signal from the detectable binding complex, thereby determining the presence of MxA at the detection zone.
13 . The method of claim 1 , further comprising:
(i) exposing the MxA to a fluorescent or phosphorescent detection reagent to form a detectable binding complex comprising the MxA coupled to the fluorescent or the phosphorescent detection reagent; and
(ii) detecting a fluorescent signal or phosphorescent signal from the detectable binding complex, thereby determining the presence of MxA at the detection zone.