Enzyme-linked immunosorbent assay for the detection of the coronavirus-like agent and its antibodies in pigs with porcine epidemic diarrhea

Callebaut, P.; Debouck, P.; Pensaert, Maurice

doi:10.1016/0378-1135(82)90009-8

Cited by 39 publications

(41 citation statements)

References 17 publications

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“…17,18 In another study in which a blocking ELISA with polyclonal antibodies for antigen capture and detection was used, PEDV was detected only in 30% of the fecal samples taken between 6 and 8 days after experimental inoculation with the CV-777 strain. 2 This finding confirms that the blocking ELISA using MAb CVI-PEDV 66.31 and CVI- PEDV 66.49 has a higher sensitivity for the detection of viral antigen.…”

Section: Discussionsupporting

confidence: 77%

“…Because until only recently the virus could not be isolated in cell culture, the development of diagnostic methods for detection of the virus or its antibodies has been impaired. Previous tests for antibody detection used viral antigen produced in experimental animals 2,5 or gut cryostat sections of infected pigs 5,15 PEDV was isolated for the first time in 1988 using the Vero cell line and a trypsin-supplemented medium. 9 Since then, different diagnostic methods 10,11,19 have been described for the detection of both PEDV antigen and its antibodies in experimentally induced infections or in a limited number of field outbreaks.…”

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confidence: 99%

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Evaluation of a Blocking ELISA Using Monoclonal Antibodies for the Detection of Porcine Epidemic Diarrhea Virus and Its Antibodies

et al. 1995

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Abstract. Two blocking enzyme-linked immunosorbent assays (ELISAs) involving the use of monoclonal antibodies (MAb) as capture and detecting agents and an indirect fluorescence test (IFT) were used for the detection of porcine epidemic diarrhea virus (PEDV) antigen or its antibodies. In the ELISA for viral antigen detection, the blocking step is accomplished by incubation of fecal samples, in duplicate wells, with PEDVspecific positive or negative serum, whereas in the ELISA for antibody detection the blocking step is accomplished by incubation of serum samples with a gut-origin virus suspension. All the methods developed were used to monitor an experimental infection in piglets with the CV-777 strain of PEDV and a natural PED outbreak in a swine fattening unit. The antigen-detection ELISA was able to detect PEDV shedding for a longer time than have previously described methods. The blocking ELISA for antibody detection was able to detect the serum antibody response sooner after PEDV infection than did IFT.

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Section: Discussionsupporting

confidence: 77%

mentioning

confidence: 99%

Evaluation of a Blocking ELISA Using Monoclonal Antibodies for the Detection of Porcine Epidemic Diarrhea Virus and Its Antibodies

et al. 1995

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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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“…Immunofluoresence and ELISA tests for the detection of PEDV antigens and antibodies have been developed already [2]. Recently, the polymerase chain reaction (PCR) has come into general use for the rapid detection of microbial pathogens and clinical diagnostics because of its high specificity and sensitivity [16].…”

mentioning

confidence: 99%

“…PED has been diagnosed by detection of virus and/or its antigens in feces, or by demonstration of seroconversion, using indirect immunofluorescence test (IIFT) [7,20,21], immunofluorecence blocking test (IFBT) [27], enzymelinked immunosorbent assay (ELISA) blocking test [2] and ELISA [9]; but these methods are time-consuming and require expertise. Recently, PCR has commonly been used to detect genes of viruses, bacteria and protozoa [17,25,26], for time-saving and because of the limited availability of samples.…”

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confidence: 99%

Detection of Porcine Epidemic Diarrhea Virus Using Polymerase Chain Reaction and Comparison of the Nucleocapsid Protein Genes among Strains of the Virus.

et al. 1999

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ABSTRACT. Two pairs of PCR primers were designed to perform nested PCR targetting of a 540 bp fragment of the nucleocapsid (N) protein gene (N gene) of porcine epidemic diarrhea virus (PEDV). The N gene of PEDV was amplified with 4 PEDV strains and 11 small intestines of PEDV-infected piglets collected from 2 farms in Kagoshima prefecture, Japan. Nucleotide sequences of the PCR products from a Korean and two Japanese strains (KKN96-1 and S1) of PEDV isolated in 1993 and 1996, respectively, were almost identical. These results suggest that the PCR is an available tool for detection of PEDV from pigs in the field, and that the two Japanese strains (KKN96-1 and S1) were genetically similar to the Korean strain.-KEY WORDS: nested PCR, nucleocapsid protein gene, porcine epidemic diarrhea virus.

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Enzyme-linked immunosorbent assay for the detection of the coronavirus-like agent and its antibodies in pigs with porcine epidemic diarrhea

Cited by 39 publications

References 17 publications

Evaluation of a Blocking ELISA Using Monoclonal Antibodies for the Detection of Porcine Epidemic Diarrhea Virus and Its Antibodies

Evaluation of a Blocking ELISA Using Monoclonal Antibodies for the Detection of Porcine Epidemic Diarrhea Virus and Its Antibodies

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

Detection of Porcine Epidemic Diarrhea Virus Using Polymerase Chain Reaction and Comparison of the Nucleocapsid Protein Genes among Strains of the Virus.

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