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Research Note: Development of an ELISA to distinguish between goose parvovirus infection and vaccine immunization antibodies.

ABSTRACT: Goose parvovirus (GPV) leads to a huge loss in the poultry industry, and early diagnosis is required to prevent the disease from spreading. At present, there are a variety of detection methods for GPV infection, and the ELISA method has the advantages of simple and rapid operation. However, most ELISA methods for detecting GPV can only detect the antibody level of the sample, but cannot distinguish between the GPV infection and vaccine immunization antibodies. Therefore, this study has a wider application value by establishing the detection method based on the structure and non-structural protein of the virus. The GPV non-structural (NS1) and structure (VP3) fusion proteins were used as coating antigens to establish 2 indirect ELISA methods, and the detection conditions were optimized. A series of experiments proved that the detection method has good specificity, sensitivity, and repeatability. The test results of 120 immune sera samples and 145 natural infection serum samples showed that the positive rates of immunized serum were 9.17% (NS1) and 88.33% (VP3), and the positive rates of natural infection were 88.97% (NS1) and 86.21% (VP3), which distinguish between the GPV infection and vaccine immunization antibodies. The establishment of 2 indirect ELISA methods using NS1 and VP3 proteins as inclusion antigens provides a new method for detecting GPV infection and inactivated immune antibodies, which lays a foundation for the serological diagnosis and epidemiological monitoring of GPV.

SUBMITTER: Zhang S

PROVIDER: S-EPMC7587739 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Research Note: Development of an ELISA to distinguish between goose parvovirus infection and vaccine immunization antibodies.

Zhang Shuai S Yang Jing J Wang Zhenzhong Z Chen Lin L Diao Youxiang Y Tang Yi Y

Poultry science 20200125 3

Goose parvovirus (GPV) leads to a huge loss in the poultry industry, and early diagnosis is required to prevent the disease from spreading. At present, there are a variety of detection methods for GPV infection, and the ELISA method has the advantages of simple and rapid operation. However, most ELISA methods for detecting GPV can only detect the antibody level of the sample, but cannot distinguish between the GPV infection and vaccine immunization antibodies. Therefore, this study has a wider a ...[more]

PMID: 32111309

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Project description:This study aimed to investigate whether the developmental changes in glucose metabolism were associated with insulin signaling in the middle and later stages of goose embryos. Serum and liver were sampled on embryonic day 19, 22, 25, 28, and day of hatchment, with 30 eggs at each sampling time point, and 6 replicates of 5 embryos. The embryonic growth traits, serum glucose, hormone levels, and the hepatic mRNA expressions of target genes related to glucose metabolism and insulin signaling were measured at each time point. Relative body weight, relative liver weight, and relative body length decreased linearly and quadratically from embryonic day 19 to day of hatchment, while relative yolk weight decreased linearly from embryonic day 19 to day of hatchment. Serum glucose, insulin, and free triiodothyronine levels increased linearly with increasing incubation time, while no differences were observed in serum glucagon and free thyroxine levels. The hepatic mRNA expression related to glucose catabolism (hexokinase, phosphofructokinase, and pyruvate kinase) and insulin signaling (insulin receptor, insulin receptor substrate protein, Src homology collagen protein, extracellular signal-regulated kinase, and ribosomal protein S6 kinase, 70 ku) increased quadratically from embryonic day 19 to day of hatchment. The expression of citrate synthase and isocitrate dehydrogenase mRNA decreased linearly and quadratically respectively from embryonic day 19 to day of hatchment. Serum glucose levels were positively related to serum insulin (r = 1.00) and free triiodothyronine (r = 0.90) levels, as well as the hepatic mRNA expression of insulin receptor (r = 1.00), insulin receptor substrate protein (r = 0.64), extracellular signal-regulated kinase (r = 0.81), and ribosomal protein S6 kinase, 70 ku (r = 0.81) related to insulin signaling. In conclusion, glucose catabolism was enhanced and had positive correlations with the insulin signaling in the middle and later stages of geese embryogenesis.

Project description:BackgroundClassic goose parvovirus (cGPV) causes high mortality and morbidity in goslings and Muscovy ducklings. Novel GPV (N-GPV) causes short beak and dwarfism syndrome (SBDS) in Cherry Valley ducks, Pekin ducks and Mule ducks. Both cGPV and N-GPV have relatively strict host specificity, with obvious differences in pathogenicity. Specific detection of cGPV and N-GPV may result in false positives due to high nucleotide similarity with Muscovy duck parvovirus (MDPV). The aim of this study was to develop a highly specific, sensitive, and reliable TaqMan real-time PCR (TaqMan qPCR) assay for facilitating the molecular detection of cGPV and N-GPV.ResultsAfter genetic comparison, the specific conserved region (located on the NS gene) of cGPV and N-GPV was selected for primer and probe design. The selected regions were significantly different from MDPV. Through a series of optimization experiments, the limit of detection was 50.2 copies/μl. The assay was highly specific for the detection of cGPV and N-GPV and no cross-reactivity was observed with E. coli., P.M., R.A., S.S., MDPV, N-MDPV, DAdV-A, DEV, GHPV, DHAV-1, DHAV-3, ATmV, AIV, MDRV and N-DRV. The assay was reproducible with an intra-assay and inter-assay variability of less than 2.37%. Combined with host specificity, the developed TaqMan qPCR can be used for cGPV and N-GPV in differential diagnoses. The frequency of cGPV in Muscovy duckling and goslings was determined to be 12 to 44%, while N-GPV frequency in Mule ducks and Cherry Valley ducks was 36 to 56%. Additionally, fluorescence-positive signals can be found in Mule duck embryos and newly hatched Mule ducklings. These findings provide evidence of possible vertical transmission of N-GPV from breeding Mule ducks to ducklings.ConclusionsWe established a quantitative platform for epidemiological investigations and pathogenesis studies of cGPV and N-GPV DNA that was highly sensitive, specific, and reproducible. N-GPV and cGPV infections can be distinguished based on host specificity.

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Research Note: Development of an ELISA to distinguish between goose parvovirus infection and vaccine immunization antibodies.

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Research Note: Development of an ELISA to distinguish between goose parvovirus infection and vaccine immunization antibodies.

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