Practical Virology Lab. (4A) Immunocytochemistry & Immunohistochemistry

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.

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.

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.

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

Immunohistochemistry IHC Immunohistochemistry

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

Principle of IHC 6 5 4 2nd Ab mAb Ag

Procedure of IHC Oven 1 Hour 3 1 2 Monoclonal Ab Xylene Oven 65ºC 99% Incubator 37ºC 95% 70% 1 Hour Tissue DW

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

HCMV in human placenta

HCMV in human placenta

HCMV in human placenta

VCAM-1

CD31 (PECAM-1)

PROGESTERONE