Method for detecting the presence of hemoglobin in a biological sample

A method for the detection of hemoglobin in a biological sample, comprising the steps of:

• (i) providing a carrier matrix which comprises a sample application region, a first and a second substrate region, and a first and a second indicator region, wherein a biological sample has been applied to the sample application region;
• (ii) contacting the first substrate region with the biological sample, wherein the first substrate region in liquid-conductive communication with, or combined with, the sample application region, wherein a pseudoperoxidase substrate comprising a peroxidase or hydroperoxidase reagent is contacted with the sample;
• and wherein the pseudoperoxidase substrate is present in the first substrate region in a dried state prior to contacting with the biological sample;
• (iii) contacting the sample and pseudoperoxidase substrate with the first indicator region, wherein the first indicator region in liquid-conductive communication with, or combined with, the first substrate region and having an indicator applied thereto or impregnated therein, wherein the sample and substrate are contacted with the indicator which produces a detectable response in the presence of heme and the pseudoperoxidase substrate;
• (iv) contacting a second substrate region with the biological sample, wherein the second substrate region is in liquid-conductive communication with the sample application region and having a detectable anti-globin immunointeractive molecule applied thereto or impregnated therein, wherein the sample is contacted with a detectable anti-globin immunointeractive molecule to form a detectable globin-antiglobin complex in the presence of globin;
• and
• (v) contacting the detectable globin-antiglobin complex with a second indicator region, wherein the second indicator region is in liquid-conductive communication with the second substrate region and having an anti-globin immunointeractive molecule immobilized therein, wherein the detectable globin-antiglobin complex is contacted with an immobilized anti-globin immunointeractive molecule to immobilize the detectable globin-antiglobin complex.