A comparison of three avidin-biotin complex immunoenzyme systems for detection of African swine fever virus antigen in paraffin-embedded tissues.

摘要: The sensitivity and specificity of 3 avidin-biotin complex (ABC) immunostaining systems were compared on paraffin-embedded tissues from African swine fever virus (ASFV)-infected pigs. Results also with immunofluorescent detection cryosections the same tissue for optimal ASFV antigen. ABC-alkaline phosphatase (ABC-AP) ABC-peroxidase (ABC-PO) at least as sensitive direct fluorescent antibody (FA) 10-fold more than ABC-glucose oxidase system. Three ABC-AP 2 ABC-PO chromagens different counterstains compared. In addition, fixatives, biotinylation procedures, 7 endogenous peroxidase blocking regimes, 6 adhesives, mounting media system a red chromagen hematoxylin counterstaining was preferred most closely approximated routinely stained pathologic sections. Fixation in paraformaldeh yde- lysine-periodate fixative preserved antigen research studies years. Formalin-fixed retained some staining up to 10 technique localization sections has been mainstay microbiologic diagnostics decades. An FA procedure (ASFV) developed 1966, 11 methods have changed little since then. Using this technique, pathogenesis elucidated, cells mononuclear phagocytic shown be primarily infected. Recent 17 using histopathology found monocytic paracortex mandibular lymph node earliest Further characterization these hampered by inherent limitations immunostaining, including poor morphology de- tection paraffin These problems limited usefulness further cells. immunoenzyme provides higher amplification other im- munostaining systems. Early uses ABC immunoglobulins, 12,19 cell mark- ers 8,10,13 hormones. 2-5,15 antigens are often abundant relatively stable viral antigens. We an