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At least 235 records · Page 13Linked to original sources
PubMed2006

Generating vesicular stomatitis virus pseudotype bearing the severe acute respiratory syndrome coronavirus spike envelope glycoprotein for rapid and safe neutralization test or cell-entry assay.

We generated a recombinant vesicular stomatitis virus (VSV) pseudotype (VSV Delta G*SG) by replacing the envelope G gene with the GFP gene and complementing with spike glycoprotein (S) of SARS-CoV in trans. The neutralization and infection blocking tests showed that the VSV Delta G*SG and SARS-CoV reacted similarly to SARS-CoV specific antiserum, suggesting the VSVDelta G*SG can be a safe replacement of the live SARS-CoV for neutralization test and cell-entry assay.

Animals↗
PubMed1995

Comparison of commercial enzyme immunoassay kits with plaque reduction neutralization test for detection of measles virus antibody.

Four commercially available enzyme immunoassay (EIA) kits were evaluated in comparison with the plaque reduction neutralization (PRN) test for detection of measles virus antibody. The EIA kits, Enzygnost (Behring), Diamedix, Vidas (bioMerieux Vitek), and Measlestat (Biowhittaker), were assessed with two PRN cutoff titers: a PRN titer of 8, the lowest detectable antibody level by the PRN test under the test conditions, and a titer of 120, which has been shown to be the minimum protective antibody titer. At a PRN cutoff titer of 8, the sensitivity was 88.2, 91.1, 74.6, and 69.8% for Behring, Diamedix, Vidas, and Biowhittaker EIA tests, respectively, with negative predictive values ranging from 22.7 to 45.5%. The specificity was 93.8% for Diamedix and 100% for the rest. At a PRN cutoff titer of 120, the sensitivity and specificity, respectively, were 100 and 90.7% (Behring), 98.2 and 58.8% (Diamedix), 90.6 and 94.5% (Vidas), and 85.7 and 96.4% (Biowhittaker). At this PRN cutoff titer, the negative predictive values of all EIA tests improved considerably, ranging from 70.7 to 100%. The EIA results showed an excellent association with PRN results when the PRN titers of the test samples were either < 8 or > 1,052. Discrepancies occurred especially when testing samples having PRN titers in the range of 8 to 120, indicating lack of sensitivity of the EIA tests in detecting measles virus antibody at low levels. Maternally derived measles virus antibody at this level has been shown to interfere with measles vaccine response in children and hence has implications from the standpoint of measles immunization. The ready availability, ease of operation, and rapid turnaround time are strong plus points of the EIA kits, and they could be useful in a clinical laboratory setting for routine application, but they may have limited use in vaccine-related studies and seroepidemiological surveys.

Antibodies, Viral↗
PubMed1983

Enzyme-linked immunosorbent assay for serum antibody to bovine respiratory syncytial virus: comparison with complement-fixation and neutralization tests.

An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of serum antibody to bovine respiratory syncytial virus (BRSV) was developed and compared with complement-fixation (CF) and viral neutralization (VN) tests. Tissue culture-grown viral antigens were used in the tests. Serum samples from various sources were compared, including serum samples from 10 calves which were infected experimentally with BRSV by aerosol exposure. The ELISA compared favorably with the VN test for detecting serologic responses and for detecting passively acquired antibody in young calves. Approximately 98% (109/111) of field serum samples which were positive by the VN test were also positive by ELISA. The ELISA and the CF test both failed to detect early appearing (1 to 4 weeks after exposure) antibody in 2 experimental calves with subclinical infection with BRSV. The CF test appeared to be less specific in that it gave positive reactions with some sera which were negative by both the ELISA and the VN test. The CF test did not detect antibody in 50% of serum samples obtained at the farm from young feeder calves which were positive by both ELISA and the VN test. The ELISA appears to be a sensitive and specific serologic procedure for detecting serum antibody to BRSV and has the advantage of giving test results within several hours, whereas the VN test requires 5 to 6 days for completion.

Animals↗
PubMed1984

Evaluation of focus reduction neutralization test with peroxidase-antiperoxidase staining technique for hemorrhagic fever with renal syndrome virus.

Titers of hemorrhagic fever with renal syndrome virus were estimated by counting foci stained with peroxidase-antiperoxidase or by using immunofluorescence methods. Foci stained with PAP were clearer and easier to count. The peroxidase-antiperoxidase method showed a linear relationship between virus concentration and the number of foci. A focus-reduction neutralizing antibody test was performed with immune rat sera and sera from patients. Again, the peroxidase-antiperoxidase method proved to be more convenient and reliable than the immunofluorescence method. Antigenic differences between Hantaan virus, 76-118 and hemorrhagic fever with renal syndrome virus B-1 were clearly demonstrated by the neutralizing antibody test.

Animals↗
PubMed2005

Comparison of an enzyme-linked immunosorbent assay with serum neutralization test for serodiagnosis of porcine epidemic diarrhea virus infection.

An indirect porcine epidemic diarrhea (PED) virus (PEDV) enzyme-linked immunosorbent assay (ELISA) was compared with the serum neutralization (SN) test by testing 46 samples from experimentally infected sows, 73 samples from naive sows, and 1,024 field sow samples from 48 commercial swine farms of undefined PED status. The SN test and the ELISA were performed using PEDV, KPEDV-9 strain. Viral proteins as a coating antigen of PEDV ELISA were extracted from the cytoplasm of PEDV-infected Vero cells using a non-ionic detergent, Triton X-100, and a simple protocol of PEDV ELISA was followed. The presence of antibodies in these experimental samples was confirmed by SN and ELISA in which the sensitivity of the ELISA was 89.1%, and the corresponding specificity was 94.5%. On testing 1,024 field samples, an overall agreement of 84.2% was generated between the SN and ELISA. This study demonstrates that the PEDV ELISA is a useful serodiagnostic screening test at herd level for detecting swine antibodies against PEDV.

Animals↗
PubMed2003

Establishment of an analyzing method for a Japanese encephalitis virus neutralization test in Vero cells.

We established a 50% plaque reduction analyzing method of neutralizing antibody for human serum to Japanese encephalitis virus (JEV) in Vero cells, called the '3 points least-squares regression method' (3LSRM). Our method shows a high correlation with the chick embryo cell method (the current standard method for human serum), using the chart method established by the National Institute of Infectious Diseases of Japan, which is an equation made with retrospective data obtained with the 50% plaque reduction method as a standard measurement method for neutralizing antibody titers to JEV. Our new method is much simpler and more reliable than the current method in that it uses an established cell line, Vero cells, and its results are computed by the 3LSRM.

Animals↗
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↗