An Enzyme-Linked Immunosorbent Assay That Measures Protective Antibody Levels to Newcastle Disease Virus in Chickens

1984 ◽  
Vol 28 (4) ◽  
pp. 1079 ◽  
Author(s):  
Richard A. Wilson ◽  
Charles Perrotta ◽  
Betsy Frey ◽  
Robert J. Eckroade
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Immunosorbent Assay ◽  
Newcastle Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protective Antibody ◽  
Antibody Levels

scholarly journals Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

2020 ◽  
Author(s):  
Pinpin Ji ◽  
Jiahong Zhu ◽  
Xiaoxuan Li ◽  
Wenqi Fan ◽  
Qianqian Liu ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Immunosorbent Assay ◽  
Newcastle Disease ◽  
Tertiary Structure ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Permanent Storage ◽  
Capture Antibody ◽  
The Cost

Abstract Background: Traditional sandwich enzyme-linked immunosorbent assay (ELISA) using polyclonal and monoclonal antibodies as reagents presents several drawbacks, including limited amounts, difficulty in permanent storage, and required use of a secondary antibody. Nanobodies can be easily expressed with different systems and fused with several tags in their tertiary structure by recombinant technology, thus offering an effective detection method for diagnostic purposes. Recently, the fenobody (ferritin-fused nanobody) and RANbody (nanobody-fused reporter) have been designed and derived from the nanobody for developing the diagnostic immunoassays. However, there was no report about developing the sandwich ELISA using the fenobody and RANbody as pairing reagents.Results: A platform for developing a sandwich ELISA utilizing fenobody as the capture antibody and RANbody as the detection antibody was firstly designed in the study. Newcastle disease virus (NDV) was selected as the antigen, from which 13 NDV-specific nanobodies were screened from an immunized Bactrian camel. Then, 5 nanobodies were selected to produce fenobodies and RANbodies. The best pairing of fenobodies (NDV-fenobody-4, 800 ng/well) and RANbodies (NDV-RANbody-49, 1:10) was determined to develop the sandwich ELISA for detecting NDV. The detection limits of the assay were determined to be 22 of hemagglutination (HA) titers and 10 ng of purified NDV particles. Compared with two commercial assays, the developed assay shows higher sensitivity and specificity. Meanwhile, it exhibits 98.7% agreement with the HA test and can detect the reference NDV strains belonging to Class II but not Class I. Conclusions: In the presented study, the 13 anti-NDV nanobodies binding the NDV particles were first produced. Then, for the first time, the sandwich ELISA to detect the NDV in the different samples has been developed using the fenobody and RANbody as reagents derived from the nanobodies. Considering the rapidly increasing generation of nanobodies, the platform can reduce the cost of production for the sandwich ELISA and be universally used to develop assays for detecting other antigens.

scholarly journals Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

2020 ◽  
Author(s):  
Pinpin Ji ◽  
Jiahong Zhu ◽  
Xiaoxuan Li ◽  
Wenqi Fan ◽  
Qianqian Liu ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Immunosorbent Assay ◽  
Newcastle Disease ◽  
Tertiary Structure ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Permanent Storage ◽  
Capture Antibody ◽  
The Cost

Abstract Background Traditional sandwich enzyme-linked immunosorbent assay (ELISA) using polyclonal and monoclonal antibodies as reagents presents several drawbacks, including limited amounts, difficulty in permanent storage, and required use of a secondary antibody. Nanobodies can be easily expressed with different systems and fused with several tags in their tertiary structure by recombinant technology, thus offering an effective detection method for diagnostic purposes. Recently, the fenobody (ferritin-fused nanobody) and RANbody (nanobody-fused reporter) have been designed and derived from the nanobody for developing the diagnostic immunoassays. However, there was no report about developing the sandwich ELISA using the fenobody and RANbody as pairing reagents. Results A platform for developing a sandwich ELISA utilizing fenobody as the capture antibody and RANbody as the detection antibody was firstly designed in the study. Newcastle disease virus (NDV) was selected as the antigen, from which 13 NDV-specific nanobodies were screened from an immunized Bactrian camel. Then, 5 nanobodies were selected to produce fenobodies and RANbodies. The best pairing of fenobodies (NDV-fenobody-4, 800 ng/well) and RANbodies (NDV-RANbody-49, 1:10) was determined to develop the sandwich ELISA for detecting NDV. The detection limits of the assay were determined to be 2 2 of hemagglutination (HA) titers and 10 ng of purified NDV particles. Compared with two commercial assays, the developed assay shows higher sensitivity and specificity. Meanwhile, it exhibits 98.7% agreement with the HA test and can detect the reference NDV strains belonging to Class II but not Class I. Conclusions In the presented study, the 13 anti-NDV nanobodies binding the NDV particles were first produced. Then, for the first time, the sandwich ELISA to detect the NDV in the different samples has been developed using the fenobody and RANbody as reagents derived from the nanobodies. Considering the rapidly increasing generation of nanobodies, the platform can reduce the cost of production for the sandwich ELISA and be universally used to develop assays for detecting other antigens.

scholarly journals Effect of Immunosuppression on Newcastle Disease Virus Persistence in Ducks with Different Immune Status

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Lucy W. Njagi ◽  
Phillip N. Nyaga ◽  
Lilly C. Bebora ◽  
Paul G. Mbuthia ◽  
Uswege M. Minga
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Immune Status ◽  
Experimental Period ◽  
Hemagglutination Inhibition Test ◽  
Viral Excretion ◽  
Antibody Titres ◽  
Antibody Levels ◽  
Chicken Embryo Fibroblasts

This study was carried out to verify the possibility that ducks are sources of Newcastle disease (ND) virus infection for chickens in mixed flocks. Immunosuppressed (IS) and non immunosuppressed (NIS) birds, at three different antibody levels (medium, low and absent) were used; the titres having been induced through vaccination, and Immunosuppression done using dexamethazone. Each of the 3 respective groups was further divided into 2 groups of about 12 ducks each: one challenged with velogenic ND virus; the other not challenged. Selected ducks from all groups had their antibody titres monitored serially using hemagglutination inhibition test, while two birds from each of the challenged groups were killed and respective tissues processed for ND viral recovery, using chicken embryo fibroblasts. In general, antibody titres of IS and NIS challenged ducks were significantly higher than their unchallenged counterparts (P<0.05). Non-challenged pre-immunised ducks had a progressive decrease in antibody levels; non-immunised ducks did not seroconvert. Newcastle disease virus was isolated from livers and kidneys of the challenged ducks throughout the experimental period; indicating a possibility of viral excretion, especially when the birds are stressed. It, therefore, provides another possible model of viral circulation within mixed flocks.

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