LYSIS IN A FLUIDIC CARTRIDGE BY MAGNETIC AGITATION OF LYSIS ELEMENTS AND WITH INTERNAL CONTROL
A fluid sample is dispensed into a sample chamber of a fluidic cartridge, the sample chamber containing non-magnetic beads and a magnetic element. A portion of the non-magnetic beads and magnetic element have an internal control reagent containing an internal control deposited thereon for validating an assay result and/or to validate the effectiveness of a cell lysis procedure. The magnetic element is exposed to a magnetic field, thereby causing movement of the magnetic element, which causes movement of the non-magnetic beads. Movement of the non-magnetic beads causes cells contained within the fluid sample to lyse and release nucleic acids. The internal control reagent dissolves in the presence of the fluid sample, thereby releasing the internal control into the fluid sample, and the movement of the magnetic element and the non-magnetic beads causes the internal control to be distributed within the fluid sample.
1 . A method for lysing cells contained in a fluid sample, comprising:
(A) dispensing the fluid sample into a sample chamber of a fluidic cartridge, the sample chamber containing a plurality of non-magnetic beads and at least one magnetic element, wherein at least a portion of the plurality of non-magnetic beads and/or the at least one magnetic element have an internal control reagent deposited thereon, and wherein an internal control contained in the internal control reagent is provided to validate an assay result and/or to validate the effectiveness of a cell lysis procedure; and
(B) after (A), subjecting the fluid sample to the cell lysis procedure, the cell lysis procedure comprising exposing the at least one magnetic element to a magnetic field, thereby causing movement of the at least one magnetic element contained within the sample chamber, the movement of the at least one magnetic element causing movement of the plurality of non-magnetic beads contained within the sample chamber, and the movement of the plurality of non-magnetic beads within the sample chamber causing cells contained within the fluid sample to lyse and release nucleic acids, wherein the internal control reagent dissolves in the presence of the fluid sample, thereby releasing the internal control into the fluid sample, and wherein the movement of the at least one magnetic element and the plurality of non-magnetic beads causes the internal control contained within the dissolved internal control reagent to be distributed within the fluid sample.
2 . The method of claim 1 , wherein the plurality of non-magnetic beads and the at least one magnetic element are contained within a hollow body defining a lysis chamber disposed within the sample chamber during (A) and (B), and wherein the receptacle is liquid permeable.
3 . The method of claim 1 , wherein each of the plurality of non-magnetic beads has a spherical shape, and wherein each of the plurality of non-magnetic beads has diameter of 100 μm to 2000 μm.
4 . The method of claim 1 , wherein the at least one magnetic element is plated or encapsulated with a non-magnetic material.
5 . The method of claim 1 , wherein the at least one magnetic element occupies a greater volume than any of the plurality of non-magnetic beads.
6 . The method of claim 1 , wherein the at least one magnetic element has the shape of a cube.
7 . The method of claim 1 , wherein the at least one magnetic element and each of the plurality of non-magnetic beads are inert.
8 . The method of claim 1 , wherein the plurality of non-magnetic beads occupies a volume of 50% to 70% of the volume of the sample chamber.
9 . The method of claim 8 , wherein the at least one magnetic element occupies a volume of 4.5% to 11% of the volume of the sample chamber.
10 . The method of claim 1 , wherein the magnetic field is created by an electromagnet during (B).
11 . The method of claim 10 , wherein (B) comprises alternating a current to the electromagnet to alternate a polarity of the electromagnet.
12 . The method of claim 11 , wherein (B) comprises alternating the current at a frequency of 20 Hertz to 200 Hertz.
13 . The method of claim 1 , further comprising:
(C) after (B), transporting at least a portion of the fluid sample from the sample chamber to a processing chamber of the fluidic cartridge.
14 . The method of claim 13 , further comprising:
(D) during (C), retaining lysed cellular material from (B) within the sample chamber while the released nucleic acids is transported to the processing chamber.
15 . The method of claim 14 , further comprising:
(E) in the processing chamber, immobilizing at least a portion of the released nucleic acids on a solid support and removing non-immobilized components of the fluid sample to a waste chamber of the fluidic cartridge.
16 . The method of claim 15 , further comprising:
(F) after (E), eluting the immobilized nucleic acids from the solid support and transporting the eluted nucleic acids to a reaction chamber of the fluidic cartridge.
17 . The method of claim 16 , further comprising:
(G) after (F), subjecting the eluted nucleic acids to conditions of a first reaction, the first reaction providing an indication of the presence or amount of an analyte of interest.
18 . The method of claim 17 , further comprising:
(H) immobilizing nucleic acids associated with the internal control (“IC nucleic acids”) on the solid support during (E);
(I) after (H), eluting the IC nucleic acids from the solid support and transporting the eluted IC nucleic acids to the reaction chamber; and
(J) after (I), subjecting the IC nucleic acids to conditions of a second reaction, the second reaction providing an indication of the extent of lysis in (B).
19 . The method of claim 18 , wherein the conditions of the first reaction and the conditions of the second reaction are the same conditions.
20 . The method of claim 19 , wherein each of the first and second reactions is a nucleic acid amplification reaction.