serial dilutions<\/a>\u00a0with\u00a0normal saline\u00a0and observed for\u00a0agglutination\u00a0(clumping).<\/span><\/p>\n\n\n\nCoating of Latex (For detection of antibodies)<\/strong><\/span><\/p>\n\n\n\n\n- To 20 \u03bcl of latex beads taken in a 1.5 ml vial add 40 \u03bcl of glycine-saline buffer.<\/span><\/li>\n\n\n\n
- Add 60 \u03bcl of antigen to the latex and incubate at 37oC for 2 hours.<\/span><\/li>\n\n\n\n
- Spin down at 5000 rpm for 10 minutes and carefully aspirate the supernatant.<\/span><\/li>\n\n\n\n
- Resuspend the pellet in 1 ml of blocking buffer and spin down at 5000 rpm for 10 minutes.<\/span><\/li>\n\n\n\n
- Repeat the washing once more.<\/span><\/li>\n\n\n\n
- Add 90 \u03bcl of blocking buffer to the pellet, mix well.<\/span><\/li>\n\n\n\n
- Incubate at 4oC, overnight.<\/span><\/li>\n<\/ol>\n\n\n\n
Agglutination Test <\/strong><\/span><\/p>\n\n\n\n\n- To 200 \u03bcl of glycine-saline buffer taken in a vial, add 4 \u03bcl of test antisera. (50 times diluted).<\/span><\/li>\n\n\n\n
- Add 50 \u03bcl of antigen to 50 \u03bcl of diluted antiserum in a 1.5 ml vial, mix well and incubate at room temperature for 10 minutes.<\/span><\/li>\n\n\n\n
- Pipette 10 \u03bcl of coated latex onto a glass slides.<\/span><\/li>\n\n\n\n
- Add 10 \u03bcl of diluted test antiserum to slide A.<\/span><\/li>\n\n\n\n
- Add 10 \u03bcl of antiserum mixed with antigen (from step 1) to B.<\/span><\/li>\n\n\n\n
- Add 10 \u03bcl of glycine-saline buffer to C.<\/span><\/li>\n\n\n\n
- Take a tooth pick and mix the content in each slide. Discard the tooth pick after using in one slide (take a new one for the next slide).<\/span><\/li>\n\n\n\n
- After mixing, wait for 2 minutes to observe the result.<\/span><\/li>\n<\/ol>\n\n\n\n