Immuno-Polymerase chain reaction (Immuno-PCR or IPCR)

1. Immuno-Polymerase chain reaction
(Immuno-PCR or IPCR)
Presented by
Shadman tariq sadiq
(91150000630)
PhD study
Supervised by
Proff.dr. Guven Ozdemir

2. Introduction
Immuno-PCR is an antigen detection system with the
exponential signal enhancement of a PCR , in which
the polymerase chain reaction (PCR) is used to
amplify a segment of marker DNA that has been
attached specifically to antigen–antibody complexes
this mean a combines the advantages of both
ELISA and PCR to detect antigen.
The specific antibodies are labeled with a DNA marker.
By using the amplification steps of the PCR, the
sensitivity can be better compared to the “classic”
ELISA

3. Who developed this method
??

4. This method is developed by
Sano, T., Smith, C. L. & Cantor, C. R.
in (1992)
Their research published in science journal
The name of research was
(( Immuno-PCR: very sensitive antigen detection by
means of specific antibody–DNA conjugates ))

6. The concept of I-PCR
The concept of immuno-PCR (polymerase chain reaction),
illustrated in Fig. 1, is quite simple, and is similar to (ELISA)
and radioimmunoassays (RIA).
1- In the original immuno-PCR scheme, a molecular linker, which
has a bispecific binding affinity for antibody and DNA, is used
to attach a marker DNA molecule specifically to an antigen–
antibody complex.
2-A segment of the attached marker DNA is amplified by PCR
with appropriate primers, and the resulting PCR products are
analyzed.
3- The presence of specific PCR products demonstrates that the
marker DNA molecules are attached specifically to antigen–
antibody complexes, indicating the presence of antigen.

7. www.genengnews.com

8. Fig. 1

9. Different Between ELISA and PCR
• The immuno-PCR is based on the same principle
than an ELISA:
- Capture of antigen (direct on the plate or indirect by
a capture molecule)
- Recognition of the antigen by a detection antibody
- Revelation of antigen/antibody complex
• The difference is that in the case of the immuno-
PCR, the detection of the antigen/antibody complex
is performed using a DNA reporter and not an
enzyme.
• Also immuno-PCR technology has a sensitivity
greater than ELISA .

11. • The immuno-PCR provides an ultrasensitive
technology which combines the molecular
specificity of antibodies with the sensitivity of the
PCR.
• The use of a DNA reporter allows to perform a signal
amplification step by PCR amplification, which is
impossible to achieve in the case of a ELISA system.
This brings a 10 to 1000 times sensitivity increase
when compared to a classical ELISA !
This increase in sensitivity can be extremely useful
for example in the case of highly diluted sample.

16. The main steps of an immuno-PCR assay are as follows:
1. Immobilization of antibodies specific for the protein
target to the surface of a vessel
2. Washing to remove unbound antibody
3. Addition of sample
4. Washing to remove unbound sample
5. Addition of a second specific antibody, coupled to
a DNA molecule
6. Washing to remove unbound antibody
7. DNA amplification and detection

17. Advantages of Immuno-PCR:
- Sensibility dramatically improved in comparison to
classical ELISA
- Possibility to quantify
- Reducted volumes and quantities
- No specificity lost

18. Benefits of IPCR
1- biological and biomedical sciences such as
diagnoses, toxins, cytokines, hormones..etc
2-powerful method for detecting low quantities of
protein antigens.
3-microbial diagnostic.

19. Thanks for listening