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Given an in vivo sample from an experimental infection, I would like to see if a bacterial protein is present.

I've been thinking about Western Blot using sera from animals infected with the bacteria. However, this strategy needs a positive control that requires prior gene cloning and expression.

There is maybe a shorter path.

Thanks

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I think that enzyme-linked immunosorbent assay (or ELISA) is the best way to do so, given you have an antibody against the protein to coat the sample wells with... You could use mice to create polyclonal antibodies against your protein by injecting the protein and collecting the serum and purifying it...as long as there isn't similar proteins to the bacterial one in the sample you are testing for...but to diagnose an infection you could use something like q-PCR assuming you can get the bacterial DNA from the sample (you could use typical bacterial DNA extraction techniques).

For more on ELISA go here. Here is a good animation on ELISA: http://highered.mheducation.com/sites/0072556781/student_view0/chapter33/animation_quiz_1.html

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  • $\begingroup$ You don't have to use PCR to diagnose diseases, ELISA or western blot is routinely used for this purpose. $\endgroup$ – Chris Dec 11 '14 at 7:44
  • $\begingroup$ @chris well... it depends on the disease too... $\endgroup$ – TanMath Dec 11 '14 at 19:36

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