Traditional immuneassays vs luminex - (Jun/15/2006 )

Hi,

I understand that IFN-Y ELISPOT assay and lymphoproliferative assays measure cytokine production and proliferation of specific T cells resp up on stimulation with a specific antigen.

I was wondering if these assays could be completely replaced by measuring cytokine profiles of vaccinated subjects using luminex platform. The control subjects would provide baseline cytokine levels and could be compared to vaccinated individuals.

May be I am missing an important point here and I look forward to hear your thoughts.

Thanks

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There's no really difference between using an ELISA or the luminex (except the luminex should allow you to measure multiple cytokines in the same aliquot of sample, an advantage if you have alot of different proteins to measure or a small amount of sample).

ELISpot gives you a slightly different readout to either ELISA or luminex as it tells you the amount of cytokine/protein a cell was making so you can see if all the cells responded to a small degree or if a few cells responded highly (the total amount could be similar in these 2 situations by ELISA/luminex.

Measuring intracellular cytokines by flow cytometry will give you similar results to the ELISpot I would think.

All the best,
Ceri

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Yeah Thats waht I thought. I wasnt too worried about the ELSIA. I guess in a way it would be qualitative too to see cytokines being produced by specifically stimulated cells. however, rather than wasting time collecting PBMNCs and stimulating them etc etc...a readout from luminex would not be to shabby....mighty expensive though!!

unfortunately I havent seen any papers that use this as a replacement for LTT (CD4+, IL-2) or ELISPOT (CD8+, IFN-Y) readouts.

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luminex检测需要特殊的仪器, 我不知道你们试验室有没有这种仪器?

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I think we may need someone to translate the above sentence into English.

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yup u are right. I think its chinese. I will see if I could get a chinese friend of mine to translate it

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you can translate at this website:

http://world.altavista.com/

this is AltaVista Babel Fish Translation website.

here is the translation:

"The luminex examination needs the special instrument, I did not know your test chamber does have this kind of instrument?"

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Could "test chamber" be "laboratory"? Possibly something got lost in translation?

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i assume that you are correct, that is what i took it to mean. i posted the translation as it came out of the translator.

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Thanks for the translation guys. Yes we have a luminex instrument (Bioplex from biorad). It can simultaneoulsy measure 10 or more cytokines in miniscule quantities of serum.... cost being the only limitation approx $1500-2000 per test (plate)...however, it is still comaprable to that many # of ELISAs as the sample size increases. I have tons of samples so will probably be able to justify its use.

coming back to the question...this is the conclusion I have come to...depending on the kind of information I wanted to extract...it would be better to use luminex. so if i stimulate PBMNCs in vitro using specific Ag....I will get more relevant info as compared to measuring serum cytokine levels that may be a result of other factors,exposures, stress etc.

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