Genetic and antigenic analysis of Chlamydia pecorum strains isolated from calves with diarrhea

OHTANI, Akifumi; KUBO, Masahito; SHIMODA, Hiroshi; OHYA, Kenji; IRIBE, Tadashi; OHISHI, Daiki; ENDOH, Daiji; OMATSU, Tsutomu; MIZUTANI, Tetsuya; FUKUSHI, Hideto; MAEDA, Ken

The sections were also used for immunohistochemical detection of C. pecorum antigens. After deparaffinization and rehydration of the paraffin sections, endogenous peroxidase activity was blocked by incubation in 3% H 2 O 2 for 10 min. Sections were incubated with 10% normal goat serum for 30 min and then with a rabbit anti-C. pecorum Bo/Yokohama strain antiserum [6] (1:2,000) for 30 min. Sections were rinsed three times in PBS containing 0.2% Tween 20 for 10 min and then incubated with Histofine Simple Stain MAX-PO (R) (Nichirei Biosciences, Inc., Tokyo, Japan) for 30 min. The sections were rinsed three times in PBS containing 0.2% Tween 20 for 10 min. The sections were incubated with 3,3′-diaminobenzidine tetrahydrochloride under the mi-croscope to control color development, washed in tap water for 10 min and rinsed with distilled water. The sections were counterstained with Meyer's hematoxylin.