1. Practical Virology Lab. (4A) Immunocytochemistry & Immunohistochemistry

2. Immunocytochemistry (ICC):
a highly sensitive and specific method to localize the antigen in the cells with a labeled antibody
also it is easy and rapid.

3. Principle The primary (monoclonal antibody) binds the antigen
The secondary antibody (conjugated or labeled by enzyme for example) binds to the bound primary antibody
The chromogenic substrate of the enzyme when reacts with the bound enzyme (the lablel) will yield a colored precipitate at the site of the antigen.

4. Applications
A. Antigenaemia Assay: Detection of the Ag in the blood. Example: detection of early HCMV Ag in neutrophils isolated from blood of patients with suspected acute HCMV infection.
Clinical application: patients with kidney transplantation in whom serology is less valuable in the diagnosis of acute HCMV infection, also in immuno-compromised patients, and HIV positive patients.

8. B. Urine cytology: in congenital infections such as congenital HCMV
C. Pap smear and ICC in diagnosing HPV (human papilloma vs) and serotyping.

9. Immunohistochemistry
IHC
Immunohistochemistry

10. Immunohistochemistry (IHC): a highly sensitive and specific method to localize the antigen in the tissue sections with a labeled antibody

11. Principle of IHC 6 5 4
2nd Ab
mAb Ag

12. Procedure of IHC Oven 1 Hour3 1 2
Monoclonal Ab
Xylene
Oven
65ºC
99%
Incubator
37ºC
95%
70%
1 Hour
Tissue DW

13. 2nd Ab Conjugate Substrate Counter stain
Tissue 4
2nd Ab
PBS tween 5
Conjugate 6
Substrate 7
Counter stain

14. HCMV in human placenta

15. HCMV in human placenta

16. HCMV in human placenta

17. VCAM-1

18. CD31 (PECAM-1)

19. PROGESTERONE