Optical density variation in indirect ELISA - (Jan/30/2008 )

Dear All,

I am using indirect ELISA kits(VLA, UK) for diagnosis of brucellosis in ruminants.
When I put samples in duplicate wells, sometimes ODs varies vastly.
When I am testing same serum samples twice or thrice eith the same kit, ODs may vary.
Also When I am testing the serum samples from same animal collected at different interval, ODs may vary. I mean initially It was positive and then it showed negative.

I would like to know from any experts that disease like brucellosis, what is humoral response of an animal during infection.
What type of response we can see during course of infection in brucellosis.

please clarify it


regards,

Bhavesh

---

may due to multichannel pipette.
Just wondering... when titrating, how many times do you mix by pipetting up and down in the well of the ELISA plate?



may due to mixing of the solution... do you properly mix the solution from the kit before using it?
or
the condition the serum samples ...ie the freeze and thaw of the samples



I am not an expert... I don't know

Hope this may help.

---

hi eaxactly as minnie mouse told u,

some technical problem while doing ELISA, nothing to do with the humoral response of the disease in the present situation.

pay lot of attention towards minor issues while performing ur ELISA.

gud luk

---

Hi,
Are you using the ELISA plates fresh? i.e. make sure that you are not using the plates that were already maintained for some time at 37C
Also as other's told, i don't think there is any problem in the response. Also make sure that the serum you are using is free of precipitates or clumps
hope this may help
All the best

---