Immunochromatography detection of multidrug-resistant
This invention provides a immunochromatography detection device that can detect PBP2′ produced specifically by a bacterium of multidrug-resistant staphylococcus with high sensitivity in a simple and rapid manner via immunochromatography detection to determine infection with multidrug-resistant staphylococcus , a diagnostic method using such detection device, and a diagnostic kit comprising such detection device.
1. A method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, comprising detection of PBP2′ via immunochromatography detection based on an antigen-antibody reaction,
(i) which uses an immunochromatography detection device comprising on a sheet-like solid-phase support:
(a) a sample supply site to which a sample solution is supplied, which sample solution is deduced to contain a bacterium producing PBP2′ or a solution deduced to contain PBP2′ released from the cell wall via sample pretreatment;
(b) a labeled reagent site that holds a reagent, which is a labeled antibody binding specifically to PBP2′, in a manner such that the reagent is able to spread across the solid-phase support; and
(c) a capture reagent site on which a capture reagent capable of specifically binding to and capturing a complex of PBP2′ and the labeled reagent has been immobilized, the capture reagent site comprising a sulfobetaine-type surfactant, and
(ii) which comprises supplying to the sample supply site the sample solution and then evaluating at the capture reagent site the formation of a complex of a labeled reagent/PBP2′/a capture reagent.
2. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 1 , wherein PBP2′ comes into contact with the labeled reagent at a site separated from a solid-phase support in advance, and a sample-reagent mixture comprising a sample solution deduced to contain a bacterium producing a cell-wall-synthesizing enzyme, PBP2′ or a solution deduced to contain PBP2′ released from the cell wall via sample pretreatment and the labeled reagent that is a labeled antibody binding specifically to PBP2′ is supplied to the sample supply site.
3. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 1 , wherein the labeled reagent is an insoluble carrier to which an antibody is bound.
4. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 1 , wherein an anion selected from the group consisting of a chloride ion, a bromide ion and an iodide ion is added to a sample solution deduced to contain a bacterium producing a cell-wall-synthesizing enzyme, PBP2′, or a solution deduced to contain PBP2′ released from the cell wall via sample pretreatment prior to supply of such solution to a sample supply site.
5. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 2 , wherein a cation selected from the group consisting of a potassium ion, a calcium ion and a magnesium ion is added to a sample solution deduced to contain a bacterium producing a cell-wall-synthesizing enzyme, PBP2′, or a solution deduced to contain PBP2′ released from the cell wall via sample pretreatment prior to supply of such solution to a sample supply site.
6. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 1 which comprises a step of pretreating a sample via alkaline treatment or neutralization.
7. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 6 , wherein the alkaline treatment is carried out using an aqueous solution of alkali metal hydroxide or carbonate or an aqueous solution of alkaline earth metal hydroxide or carbonate.
8. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 7 , wherein the pH of the aqueous solution of alkali metal hydroxide or carbonate or the aqueous solution of alkaline earth metal hydroxide or carbonate is 11 or higher.
9. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 7 , wherein the concentration of the alkali metal hydroxide or carbonate or alkaline earth metal hydroxide or carbonate is between 0.01 N and 1.0 N.
10. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 6 , wherein the aqueous solution after alkali treatment is neutralized with a buffer.
11. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 1 , wherein the sample supply site comprises glass fibers.
12. The method for detecting a bacterium that produces a cell-wall-synthesizing enzyme, PBP2′, according to claim 1 , wherein the bacterium producing a cell-wall-synthesizing enzyme, PBP2′, is a multidrug-resistant staphylococcus.