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PubMed1994

A modified serum neutralization test for the detection of antibody to porcine reproductive and respiratory syndrome virus in swine sera.

Various conditions were evaluated and modified to improve the sensitivity of the serum neutralization (SN) test for detecting antibody in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV). Higher SN titers were consistently obtained by the addition of 20% fresh swine serum to the virus diluent and by the use of a permissive cell clone (MARC-145) derived from the MA-104 cell line. Test sera used to assess the SN test were obtained from 2 groups of 3-week-old pigs infected intranasally with PRRSV (MN-1b). Using the modified method, SN antibody was first detected 9-11 days postinoculation (PI), with a peak evident at 11-21 days PI. The antibody subsequently declined, and a second peak was observed between 41 and 45 days PI. The first antibody peak was not observed and the SN antibody was only detectable between 32 and 41 days PI when the test was done with 20% heated swine serum or without supplemental swine serum. The SN antibody during 2-3 weeks PI was found to be sensitive to 2-mercaptoethanol or anti-swine IgM treatment. The SN antibody titers were high when homologous PRRSV isolate was used in the test but were markedly low for heterologous PRRSV isolates. No difference in antibody titers was observed when homologous and heterologous PRRSV isolates were tested by indirect fluorescent antibody assay. These results indicate that the modified SN method is useful in detecting earlier and higher PRRSV antibody and that it can differentiate among PRRSV isolates.

Animals↗
PubMed1965

SEROLOGIC STUDIES OF AFRICAN HORSE-SICKNESS VIRUS WITH EMPHASIS ON NEUTRALIZATION TEST IN TISSUE CULTURE.

In place of mice, monkey kidney stable (MS) cell cultures were used successfully in serologic studies of African horse-sickness virus.The maintenance medium containing 2% serum was chosen as the virus diluent. Maximum neutralization occurred after 1-hour incubation at 37 C., and maintained the same titer during an additional 4-hour incubation period. No significant difference was observed between neutralization titers titrated using the same antiserum mixed with two different passage levels of virus. Rabbit and guinea pig antiserums prepared using virus grown in MS cell cultures had antibody titer as high as those prepared in the same manner using mouse brain suspension. African horse-sickness virus strains isolated in Asia were serologically identified using a standard neutralization technique in tissue culture. All the strains were closely related to each other and all had antigenic similarity to Type 6 virus (strain 114).

African Horse Sickness Virus↗
PubMedDate not supplied

Comparison of enzyme-linked immunosorbent assays and virus neutralization test for detection of antibodies to avian pneumovirus.

Two different whole-virus enzyme-linked immunosorbent assays (ELISAs), developed in Ohio (OH) with APV/Minnesota/turkey/2a/97 and in Minnesota (MN) with APV/Colorado/turkey/97, and the virus neutralization (VN) test were used to test 270 turkey serum samples from 27 Minnesota turkey flocks for avian pneumovirus (APV) antibodies. In addition, 77 turkey serum samples and 128 ostrich serum samples from Ohio were tested. None of the turkey samples from Ohio had antibodies to APV by the VN test and OH ELISA. The ostrich samples were only tested with the VN test and were all negative for antibodies to APV. For the Minnesota serum samples, 107, 115, and 120 were positive by the VN test, the OH ELISA, and the MN ELISA, respectively. The Kappa values of 0.938 and 0.825 showed excellent agreement between the VN test and the OH ELISA and the MN ELISA, respectively, for detection of antibodies to the APV. The OH ELISA and MN ELISA had sensitivities of 1.0 and 0.953, specificities of 0.950 and 0.889, and accuracies of 0.970 and 0.914, respectively. Our results indicate that the 3 methods are sensitive and specific for diagnosis of the APV infection.

Animals↗