THE DEVELOPMENT OF IMMUNOCHROMATOGRAPHIC DIAGNOSTIC KIT THAT CAPABLE OF USING WHOLE BLOOD AS SAMPLE

• Main Author: Jatiningsih, Rahayu
• Format: Theses
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/47665
• Institution: Institut Teknologi Bandung

Immunochromatographic based diagnostic kit is one of many types of diagnostic kits and the kit commonly uses blood serum or plasma as sample for analytes detection. Whole blood can not be used as blood cells will reduce kit performance or disrupt the kit. The use of serum or plasma as sample however has weakness as the preparation of these samples require centrifugation and centrifuge does not always available at Indonesian health center. Agglutinin is a group of substance known capable of forming agglutinations of red blood cells. Spesific antibody that produced by immunizations of specific antigen known has ability as agglutinin. The performance of agglutinin binding to red blood cells is expected avoid disruption of red blood cells in the performance of Immunochromatographic based diagnostic kit. Agglutinations require anticoagulant and Tripsynization in red blood cells for good result. However, trypsinization give less effectiveness of the development immunochromatographic based diagnostic kit that capable of using whole blood as sample. Based on that, the aim of purpose of this research is to develop immunochromatographic diagnostic kit with agglutinin (crude extract of IgY anti human red blood cells), selected anticoagulant, and without blood trypsinization as component that enable the kit to use. Laying hens divided into two groups red blood cells immunizations, that is intramuscular and subcutaneous. Immunizations takes four times by two weeks range until eight weeks. Eggs collection did after last immunizations (eight weeks), then isolate the crude extract IgY anti human red blood cells fom the eggs. Total IgY and the purity of IgY have been measured. Haemagglutinations test, agglutinations test using microscopic, anticoagulant comparison test, blood in and without trypsinized test, and confirmation test using immunochromatoghraphic based diagnostic kit have been done. The result shows that immunization route does not affect IgY total concentration in yolk, but intramuscular route immunization give better immune response than subcutaneous immunization route (P<0,05). Both Sodium citrate dextrose (Alsever) and Heparine can used as anticoagulant and does not interfere crude extract IgY anti human red blood cells to agglutinate red blood cells. Furthermore, also found that the crude extract of IgY anti human red blood cells can still agglutinate red blood cells without trypsinization.. Brightness intensity (Grey value) in nitrocellulose membrane of diagnostic test that include whole blood, crude extract IgY anti human red blood cells, and alsever’s anticoagulant as component sample, give value 178,39. There is no significant difference with blood plasma as positive control with value 188,54 (P>0,05). The use of the crude extract of IgY anti human red blood cells, anticoagulant, and without blood trypsinization as component of immunochromatoghrapic diagnostic kit, enable the kit to use whole blood as sample.