SN/T 1395.2-2005Avian Chlamydia agar immunodiffusion test for (English PDF)

3.1 Equipment

3.1.1 Electrothermal constant temperature incubator (37 ° C).

3.1.2 Microwave oven.

3.1.3 Electronic balance with an accuracy of 0.01g.

3.1.4 Observe the light on a dark background.

3.1.5 Punch. 7 holes and 1 set of plum-shaped metal puncher, aperture 5mm, hole spacing 3mm.

3.2 Reagents and materials

3.2.1 Distilled water

Meet the requirements of the third grade water in GB/T 6682.

3.2.2 Antigen, standard positive serum

Use according to the instructions.

3.2.3 Tested serum

According to the routine aseptic blood collection and separation of serum, the serum to be tested should be non-hemolyzed, not spoiled, and stored at 4 °C.

3.2.4 Agarose gel

See Appendix A for preparation and Appendix B for PBS used.

4.1 Agarose plate preparation

Slowly pour the agarose gel that has been fully melted in a microwave oven into the dish. The gel has a thickness of about 3 mm and is allowed to stand.

The plate is capped and inverted and stored at 4 ° C for 2 weeks.

4.2 Agarose plate punching

Use a plum-shaped punch to punch holes on the agar plate, see Figure C in Appendix C. 1, the hole agar carefully picked with a needle or with a negative pressure Pipe aspirate, do not destroy the surrounding agar; it is best to put the bottom of the plate on the flame and lightly bake the back of the plate before loading.

4.3 Loading

Refer to Appendix C, Figure C. 2 method of loading, the middle hole plus antigen, the serum to be tested and the positive control serum are added to the surrounding holes, and the sample is added.

The amount should be filled with agar holes, but not spilled.