Rapid Diagnosis of Porcine Epidemic Diarrhea Virus Infection by Polymerase Chain Reaction.

Kweon, Chang-Hee; Lee, Jae-Gil; Han, Myung Guk; Kang, Yong-Gu

doi:10.1292/jvms.59.231

Cited by 40 publications

(34 citation statements)

References 6 publications

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“…2,3,7,23 PEDV antigen has been detected in jejunal and ileal villus enterocytes of swine by monoclonal and polyclonal antibody-based immunohistochemical procedures. 16,21,29 In situ hybridization is a valuable adjunct to standard RNA extraction techniques for evaluating gene expression in tissues and cells.…”

Section: In Situ Hybridization For the Detection And Localization Ofmentioning

confidence: 99%

In Situ Hybridization for the Detection and Localization of Porcine Epidemic Diarrhea Virus in the Intestinal Tissues from Naturally Infected Piglets

Kim

Chae

2000

Vet Pathol

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Abstract. Detection and localization of porcine epidemic diarrhea virus (PEDV) was studied by in situ hybridization with a nonradioactive digoxigenin-labeled probe in formalin-fixed, paraffin-embedded tissues from 10 naturally infected piglets. A 377-base pair cDNA probe for viral RNA encoding the membrane proteins of PEDV cell-culture-adapted strain V215/78 was generated by the reverse transcription polymerase chain reaction. In the retrospective study of pigs from herds with diarrhea, the 10 piglets naturally infected with PEDV had positive signals for PEDV by in situ hybridization. When intestinal tissues were hybridized with the PEDV probe, a strong signal was seen in the villus enterocytes of jejunum and ileum but not in the cecum and colon. Positive cells typically had dark brown reaction products in the cytoplasm. Scattered epithelial cells along the ileal Peyer's patches dome areas contained viral RNA. In one piglet, hybridization signal was also found in the duodenum. PEDV was not demonstrated in tissues outside of the intestinal tract. These findings indicate that jejunal and ileal villus enterocytes are the main target of PEDV replication during epizootic outbreaks of the disease.

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Section: In Situ Hybridization For the Detection And Localization Ofmentioning

confidence: 99%

In Situ Hybridization for the Detection and Localization of Porcine Epidemic Diarrhea Virus in the Intestinal Tissues from Naturally Infected Piglets

Kim

Chae

2000

Vet Pathol

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“…Due to the similarities in causative agents of diarrhea, the diagnosis of PED can not only depend on the clinical signs and histopathological lesions (Kweon et al 1997). In this study nested RT-PCR was used for the detection of the PEDV based on a partial amplification of the M gene.…”

Section: Discussionmentioning

confidence: 99%

Isolation and genetic analysis of a variant porcine epidemic diarrhea virus in China

Li¹,

Tian

Liu

et al. 2016

Polish Journal of Veterinary Sciences

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Porcine epidemic diarrhea virus (PEDV) is having a severe effect on the pig breeding industry in central China. The mucosa and the content of the small intestine from newborn pre-weaned piglets with diarrhea were tested for the presence of PEDV by molecular and morphologic methods, and found to be positive. Negative-staining electron microscopy (EM) revealed the presence of coronavirus-like particles in the samples. The result of molecular detection by nested RT-PCR based on the amplification of the M gene was positive. Using a novel alternative method we successfully propagated the PEDV strain (CH/QX-2) in Vero cells, confirmed by ultrathin sections of the cells and Immunofluorescence assay (IFA). Phylogenetic analysis based on the partial S gene showed that the CH/QX-2 isolate was genetically closer to strains more commonly found in China, but differed genetically from two domestic strains (CH/S, 1986 and LZC, 2007), Korean strains (DR13, 2007, and the vaccine strain (CV777 vs) currently being used in China. CH/QX-2 formed a unique clade in the derived phylogenetic tree indicating that the CH/QX-2 strain currently circulating in central China is a new variant of PEDV. This study extends current knowledge on the diversity and epidemiology of PEDV.

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“…The viral DNA was extracted 3 times with the same volume of phenol: chloroform: isoamylalcohol (25:24:1) and then 3 times with chloroform: isoamylalcohol (25:24:1). P1: 5'-GGATTGCAAACCTGAATTCTCG-3' (sequence positions 2469 to 2490 described by Collett et al [4]) and P2: 5'-GTCCGTACCACAGTTGTGGC-3' (sequence positions 2908 to 2928) were mixed with viral DNA from recombinant BHV-1, and PCR amplification was performed with PFU polymerase (Stratagene, U.S.A.) as described [13]. PCR products were then transferred to Duralon-UV membrane (Stratagene, U.S.A.).…”

Section: Cells and Virusmentioning

confidence: 99%

Bovine Herpes Virus Expressing Envelope Protein(E2) of Bovine Viral Diarrhea Virus As a Vaccine Candidate.

Kweon

Kang

Choi

et al. 1999

The Journal of Veterinary Medical Science

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ABSTRACT. The gene encoding the envelope protein (E2) of bovine viral diarrhea virus (BVDV) was expressed under the thymidine kinase (TK) promoter of Korean bovine herpesvirus 1 (BHV-1) isolate. Thymidine kinase negative (TK ) BHV-1 recombinants expressing E2 of BVDV were constructed and the expression of E2 was identified by immunofluorescence and Western blotting. Compared to wild type BHV-1, the recombinant BHV-1 had a delayed cytopathogenic effect in cells. The immunogenicity of the recombinant BHV-1 was examined in guinea pigs and cattle. Although an increase in body temperature was detected for a few days, the inoculated cattle returned to normal temperature with the development of neutralizing antibodies to BVDV.-KEY WORDS: BVDV, expression of E2 protein, TK BHV-1 recombinant, viral vector.

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Rapid Diagnosis of Porcine Epidemic Diarrhea Virus Infection by Polymerase Chain Reaction.

Cited by 40 publications

References 6 publications

In Situ Hybridization for the Detection and Localization of Porcine Epidemic Diarrhea Virus in the Intestinal Tissues from Naturally Infected Piglets

In Situ Hybridization for the Detection and Localization of Porcine Epidemic Diarrhea Virus in the Intestinal Tissues from Naturally Infected Piglets

Isolation and genetic analysis of a variant porcine epidemic diarrhea virus in China

Bovine Herpes Virus Expressing Envelope Protein(E2) of Bovine Viral Diarrhea Virus As a Vaccine Candidate.

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