Detection of Porcine Epidemic Diarrhea Virus by Immunohistochemistry with Recombinant Antibody Produced in Phages.

Abstract

. Several diagnostic methods including immunofluorescence, enzyme-linked immunosorbent assay, polymerase chain reaction and immunohistochemistry have been developed for the detection of porcine epidemic diarrhea virus (PEDV). An immunohistochemical method using a new recombinant antibody produced by a phage antibody system (PAS16) kit was investigated and compared with that using a monoclonal antibody for PEDV detection in PEDV-infected piglets. In both the immunohistochemical methods, PEDV antigens were detected in the cytoplasm of villous enterocytes and in the macrophages infiltrated in the lamina propria at 18 to 110 hr post inoculation. The positive signals with the recombinant PAS16 antibody were similar to those with the monoclonal antibody. This result suggests that the recombinant PAS16 antibody can be applicable for the rapid immunohistochemical diagnosis of PEDV infection. and the pathogen was diagnosed as PEDV by PCR [13]. The fecal samples of piglets were emulsified by 30% with phosphate buffered saline (PBS). The emulsions were centrifuged at 13,400 × g for 15 min. The supernatant was treated with antibiotics for 2 hr at 37°C and then passed through a 0.2-µm filter. Ten colostrum-deprived piglets were reared in three positive plastic isolators at 30°C. All piglets were fed with sterilized milk replaces. Seven piglets were inoculated per os with 5 ml of fecal samples of PEDV and one piglet was inoculated with KPEDV-9 (Korean strain) as a positive control. Two piglets were inoculated with PBS as negative controls. The animals were observed at regular intervals for signs of diarrhea. Piglets showing diarrhea were immediately killed by cutting the carotid blood vessels. Tissues were fixed in 10% phosphatebuffered formalin, dehydrated, embedded in paraffin wax,

Comparative Proteome Analysis of Porcine Jejunum Tissues in Response to a Virulent Strain of Porcine Epidemic Diarrhea Virus and Its Attenuated Strain

“…The small intestine, especially the jejunum, has been reported as the target tissue of PEDV infection [21]. Therefore, in the present study, iTRAQ labeling coupled with liquid chromatography tandem-mass spectrometry (LC-MS/MS) was used to analyze whole cell changes in jejunum of piglets infected with a highly virulent PEDV strain YN13 and its cell-adapted strain YN144 for a better understanding of the virus-host interaction during PEDV infection and variation in the pathogenicity of these two strains.…”

Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor

“…As noted earlier, epidemic PEDV strains are highly enteropathogenic and mainly infect villous epithelial cells of the small intestine before disseminating (11,12). It is well known that the intestinal epithelium has two important functions: absorbing helpful substances and providing a physical fence against harmful substances.…”

The spray-drying process is sufficient to inactivate infectious porcine epidemic diarrhea virus in plasma

“…Similar to the FECV infection in cats, PEDV infection in pigs appears to be restricted to intestinal cells (Jung et al, 2014;Kim et al, 1999;Stevenson et al, 2013). PEDV antigen has also been demonstrated in macrophages infiltrating the gut lamina propria (between 18 and 110 h post inoculation) (Lee et al, 2000) and more recently, in alveolar macrophages (Park and Shin, 2014). Transient detection of PEDV RNA in serum samples could be partially explained by association of PEDV with macrophages in acutely infected pigs.…”