Pretreatment of Whole Blood for Use in Immunochromatographic Assays for Hepatitis B Virus Surface Antigen

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Abstract
Immunochromatographic assays (ICAs) are also referred to as rapid tests, since they are simple and the results can be obtained within minutes after manually loading a few drops of a sample into each sample well of the test device. However, whole blood cannot be tested with ICA kits due to the visual hindrance caused by the color of red blood cells (RBCs), unless a cell-removing device such as a filter is mounted on the kits. Thus, when testing with blood, the advantage of the ICA kit is lost because of the additional time and machines required to coagulate and separate whole blood before preparing the serum. To overcome this limitation, whole-blood samples were added to a pretreatment solution to decolor the RBCs; the resulting mixtures were then loaded into the sample wells of the test device. The pretreating solution was composed of hydrogen peroxide (H 2 O 2 ) to decolor the RBCs, Sag 471 (Osi Specialties) to restrain the mixture from vigorous foaming, sodium azide (NaN 3 ) to inhibit the enzyme, which generates excessive foam at the beginning of decolorization, and EDTA as a chelating agent. As a result of this pretreatment, whole blood could be used with the ICA kit without reducing its simplicity and rapidity.