Practical Virology Lab 4 A Immunocytochemistry Immunohistochemistry Immunocytochemistry

Practical Virology Lab. (4 A) 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.

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 5 4 2 nd Ab m. Ab Ag 6

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

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

HCMV in human placenta

HCMV in human placenta

HCMV in human placenta

VCAM-1

CD 31 (PECAM-1)

PROGESTERONE