Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome(PRRS) Virus in Swine Sera by Enzyme-Linked Immunosorbent Assay.

Takikawa, Noriyasu; Kobayashi, Shigeki; Ide, Seiya; Yamane, Y; Tanaka, Yoshio; Yamagishi, H

doi:10.1292/jvms.58.355

Cited by 18 publications

(21 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The IDEXX ELISA detected the pres-ence of antibodies in serum samples from 14 to 105 dpi (11). Although results for other indirect ELISAs have been reported previously (4,9,25,29,33), the IDEXX ELISA has gained wide acceptance by the diagnostic community as a screening test because of the good repeatability, good reproducibility, fast turnaround time, and relatively low cost of the assay.…”

mentioning

confidence: 99%

Validation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus

Ferrin

Fāng

Johnson³

et al. 2004

Clin Vaccine Immunol

View full text Add to dashboard Cite

mentioning

confidence: 99%

Validation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus

Ferrin

Fāng

Johnson³

et al. 2004

Clin Vaccine Immunol

View full text Add to dashboard Cite

“…In the negative farm, the case seemed false positive in ELISA was observed, but this was a very rare case. Takikawa et al [22] reported it was observed many non-coincident cases of ELISA positive and IFA negative, and speculated these cases were false negative in IFA. In this study, it could be clear that many of such non-coincident cases were observed in PRRSV positive farms, and high rate of these cases were PRRSV positive as the judgement of ELISA.…”

Section: Discussionmentioning

confidence: 99%

“…IFA has been most frequently used for PRRS in Japan. Recently, however, the ELISA has been developed and has increasingly displaced the IFA for diagnostic assay because of its simple protocol and high sensitivity [6,11,22,30]. The judgement of the ELISA results was analyzed in gnotobiotic pigs experimentally infected with the PRRSV and a cut-off point was set at an S/P ratio of 0.4 [30].…”

mentioning

confidence: 99%

Evaluation of Enzyme-Linked Immunosorbent Assay (ELISA)and Immunofluorescent Antibody (IFA) Test for the Detection of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Antibody in Pigs from Conventional Farms.

Yahara¹,

Ohkubo²,

Kariwa

et al. 2002

J. Vet. Med. Sci.

View full text Add to dashboard Cite

ABSTRACT. To evaluate the immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for detecting the porcine reproductive and respiratory syndrome virus (PRRSV) antibody, conventional pigs in PRRSV-positive and -negative commerci al farms were examined. Antibody development patterns in ELISA and IFA tests were compared in 3 week old piglets experimentally infected with the PRRSV. The virus was detected from 2 days post infection (PI) and then the antibody titers and S/P ratios rose by both methods. A total of 208 serum samples were collected from 4 PRRSV-negative farms and 210 samples from PRRSV-positive farms, and were tested for the PRRSV antibody by IFA and ELISA. The titer of 64 should be set as the cut-off point in IFA for field sera. Similarly, the cut-off S/P ratio should be set at 0.4 in ELISA. A high degree of correlation was observed between antibody titers by the two methods in these 418 samples, with a correlation coefficient of 0.84. The coincidence rate between the two tests was 84.7% (354/418). In non-coincident cases, ELISA was able to detect the antibody with a low titer in the serum samples which were negative in IFA but from PRRSV positive farms. ELISA was more sensitive than IFA to detect PRRSV infected animals or farms.

show abstract

“…The cells were grown in Eagle's minimum essential medium (MEM) containing 8% fetal calf serum. The maintenance medium was MEM containing 2% fetal calf serum and 50 µg/ml of gentamycin.Virus: The EDRD-1 strain of PRRS virus at the 10 to 13th passage level in MARC-145 cell cultures was used [7]. Culture fluid harvested from infected MARC-145 cell cultures was clarified by centrifugation and stored at 80°C…”

mentioning

confidence: 99%

“…The infectivity assay was carried out in 96-well microplate cultures of MARC-145 cells. The inoculated cultures were examined for CPE after incubation at 37 °C for 5 days, and the 50% infectious dose (TCID 50 ) was calculated.Experimental infection of pigs: The method of experimental infection has been described previously [7]. Briefly, three 7-week-old pigs (No.…”

mentioning

confidence: 99%

Early Serodiagnosis of Porcine Reproductive and Respiratory Syndrome Virus Infection of Pigs by Detection of Slow-Reacting and Complement-Requiring Neutralizing Antibodi.

Takikawa

Kobayashi

Ide

et al. 1997

The Journal of Veterinary Medical Science

Self Cite

View full text Add to dashboard Cite

ABSTRACT. In order to evaluate and enhance the sensitivity of the neutralization (NT) test for detecting antibody in pigs infected with porcine reproductive and respiratory syndrome (PRRS) virus, the effect of altered incubation conditions and complement use on neutralizing (NT) antibody titer were investigated. Higher NT antibody titers were consistently obtained by addition of 20% guinea pig fresh serum to virus-serum mixtures in NT tests. Furthermore, the complement-requiring NT antibody titer increased in many serum samples when the virus-serum mixtures, rather than being incubated at 37 °C for 60 min, were incubated first at 4 °C for 48 hr and then with a complement at 37 °C for 60 min. The slow-reacting and complement-requiring NT antibody was detected as early as 8 days post-inoculation. It was detected in sera collected at 8 to 28 days post-inoculation and was sensitive to 2-mercaptoethanol treatment. Sera collected at 35 to 44 days post-inoculation contained 2-mercaptoethanol resistant NT antibodies. These results indicate that the modified NT test is useful for early serodiagnosis of PRRS virus infection through detection of higher NT antibody titers, and in detecting them earlier. MATERIALS AND METHODSCell culture: The MARC-145 cells were derived from a monkey kidney cell line (MA-104) [4]. The cells were grown in Eagle's minimum essential medium (MEM) containing 8% fetal calf serum. The maintenance medium was MEM containing 2% fetal calf serum and 50 µg/ml of gentamycin.Virus: The EDRD-1 strain of PRRS virus at the 10 to 13th passage level in MARC-145 cell cultures was used [7]. Culture fluid harvested from infected MARC-145 cell cultures was clarified by centrifugation and stored at 80°C. The infectivity assay was carried out in 96-well microplate cultures of MARC-145 cells. The inoculated cultures were examined for CPE after incubation at 37 °C for 5 days, and the 50% infectious dose (TCID 50 ) was calculated.Experimental infection of pigs: The method of experimental infection has been described previously [7]. Briefly, three 7-week-old pigs (No. 1-3) initially seronegative for the PRRS virus were infected intranasally with 2 ml of 10 6.4 TCID 50 /ml of the EDRD-1 strain of the PRRS virus. One pig (No. 4) served as an in-contact control. Serial serum samples were collected daily until 14 days post-inoculation, after which they were collected once weekly.NT test: The NT test was carried out in flat-bottomed 96-well microplates. In tests, serial 2-fold dilutions of the serum inactivated at 56°C for 30 min were made in the maintenance medium. In conventional NT tests, 120 µl of each dilution was mixed with an equal volume of Porcine reproductive and respiratory syndrome (PRRS) virus, family Arteriviridae, causes reproductive disturbance in sows and respiratory distress in piglets, and is prevalent worldwide [1,2,4,5,[7][8][9][10].The neutralization (NT) test, the immunoperioxidase monolayer assay (IPMA), the indirect immunofluorescence (IIF), and the enzyme-linked immunosorbent assay (ELISA) have all...

show abstract

Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome(PRRS) Virus in Swine Sera by Enzyme-Linked Immunosorbent Assay.

Cited by 18 publications

References 0 publications

Validation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus

Validation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus

Evaluation of Enzyme-Linked Immunosorbent Assay (ELISA)and Immunofluorescent Antibody (IFA) Test for the Detection of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Antibody in Pigs from Conventional Farms.

Early Serodiagnosis of Porcine Reproductive and Respiratory Syndrome Virus Infection of Pigs by Detection of Slow-Reacting and Complement-Requiring Neutralizing Antibodi.

Contact Info

Product

Resources

About