A comparison of a blocking ELISA and a haemagglutination inhibition assay for the detection of antibodies to Avibacterium (Haemophilus) paragallinarum in sera from artificially infected chickens

Sun, Hsin‐Yun; DeYuan, Miao; Zhang, P.; Gong, Yang; Blackall, P. J.

doi:10.1016/j.biologicals.2007.04.004

Cited by 4 publications

(6 citation statements)

References 11 publications

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“…The efficacy of the I-ELISA showed 100% sensitivity in all coating serovars, but the specificity of all serovars of I-ELISA was low ( approximately 30%) because of the high cross-reactivity among the three A. paragallinarum serovars. This is in contrast to previ ous studies on monoclonal antibody based blocking ELISA that reported serovar specificity (Zhang et al, 1999;Sun et al, 2007). The validation tests indicated that the new I-ELISA was highly sensitive to detect an antibody response against A. paragallinarum from immunized chickens, both of natural infection and vaccination, compared to A. paragallinarum-free chickens.…”

Section: Chapter V Discussion and Conclusioncontrasting

confidence: 56%

“…Moreover, there is no commercial test kit for HI method. The alternative method is the monoclonal antibodybased blocking enzyme-linked immunosorbent assay (blocking ELISA) (Miao et al, 2000;Sun et al, 2 0 0 7 ) but it is mainly used in research. Although blocking ELISA using monoclonal antibody shows high specificity and acceptable sensitivity (Zhang et al, 1999), there are some limitation in testing such as (1) monoclonal antibody have only for Page serovar A and C (2) monoclonal antibody are not commercial available ( 3) some isolates of A. paragallinarum do not react with monoclonal antibody (Blackall, 1999).…”

Section: Importance and Rationalementioning

confidence: 99%

“…Uncertainty of testing results depends on individual skills and method of preparation. The alternative method is monoclonal antibody-based blocking enzyme-linked immunosorbent assay (blocking ELISA) (Miao et al, 2000;Sun et al, 2007) which is highly specific with acceptable sensitivity (Zhang et al, 1999). They reported that the sensitivity of blocking ELISA against infectious coryza for serovar A and serovar C were 78.7% and 64.7%, respectively, and the specificity of blocking ELISA against infectious coryza for serovar A and C were 99.…”

Section: Serological Detectionmentioning

confidence: 99%

“…Antigen for coating plate of developed I-ELISA and using in HI test was prepared as same as previously described method (Sun et al, 2007) with some modification.…”

Section: Antigen Preparationmentioning

confidence: 99%

“…The bacterial cells were cultured in GC agar base, harvested in a centrifuge tube and washed for three times in sterile PBS. Then, antigen for coating plate of developed I-ELISA was prepared as same as previously described method (Sun et al, 2007). In addition, the total amount of bacterium which were tested by hemagglutination were about 2 7 -2 9 for stock storage before use.…”

Section: Antigen Preparationmentioning

confidence: 99%

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Development of Elisa Test Kit for Antibody Against Avibacterium Paragallinarum

Hongprasertkul

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One hundred Babcock 308 female-layer chickens were randomly divided into five groups of 20 each. Groups 1, 2, 3 and 4 were different positive control groups of immunized chickens with commercial trivalent mineral oil vaccine, and prepared bacterins of Avibacterium paragallinarum serovars A (221), B (0222) and C (Modesto), respectively. The chickens in Group 5 were assigned as a negative control and immunized with PBS. The serum from Groups 1–5 at 4 weeks after the first vaccination were used to calculate the sensitivity and specificity of the newly developed indirect enzyme-linked immunosorbent assay (I-ELISA) method. Forty negative control sera (taken before vaccination) were used to evaluate the cut-off value of the I-ELISA against each serovar of A. paragallinarum under optimal conditions. The cut-off values of serovars A, B and C, calculated by the mean optical density of all the negative sera plus three standard deviations were 0.334, 0.484 and 0.678, respectively. The efficacy of the developed I-ELISA showed 100% sensitivity for all three serovars of coating antigen but with a low specificity of 30% for all three serovars because of the high cross reactivity among serovars. Nevertheless, the serovar A I-ELISA gave a higher response to serovar A antibodies than to the other two heterologous serovars (P < 0.05). In contrast, the I-ELISA results for B and C did not show any significant difference between the homologous and heterologous serovars. In conclusion, this newly developed I-ELISA could be an alternative method for differentiating between A. paragallinarum-free chickens and those that have received either a vaccination and/or a challenge exposure.

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Section: Chapter V Discussion and Conclusioncontrasting

confidence: 56%

Section: Importance and Rationalementioning

confidence: 99%

Section: Serological Detectionmentioning

confidence: 99%

“…Antigen for coating plate of developed I-ELISA and using in HI test was prepared as same as previously described method (Sun et al, 2007) with some modification.…”

Section: Antigen Preparationmentioning

confidence: 99%

Section: Antigen Preparationmentioning

confidence: 99%

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Development of Elisa Test Kit for Antibody Against Avibacterium Paragallinarum

Hongprasertkul

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Haemophilus

Sandal

Corbeil

Inzana

2010

Pathogenesis of Bacterial Infections in Animals

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Development of an Enzyme-Linked Immunosorbent Assay for the Measurement of Antibodies Against Infectious Coryza Vaccine

et al. 2012

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Infectious coryza is an acute respiratory disease caused by infection with Avibacterium (Haemophilus) paragallinarum. It is characterized by nasal discharge and facial swelling and is associated with growth retardation and a reduction in egg production. Hemagglutination inhibition (HI) tests are used to estimate vaccine-induced immunity against infectious coryza in vitro; however, these procedures are complicated and their sensitivity is insufficient. To address these problems, an enzyme-linked immunosorbent assay (ELISA) technique using serovar-specific regions of HMTp210 (210 kDa), an outer-membrane protein of A. paragallinarum, was developed to measure the antibodies against infectious coryza. Chickens with an ELISA titer of 0.3 or more did not exhibit clinical signs of infectious coryza against challenge with A. paragallinarum, although their HI antibody titers were negative. On the other hand, chickens with an ELISA titer below 0.3 exhibited clinical signs of the disease with one exception. Antibody prevalence rates on ELISA were 80% and 60% against infection with serovars A and C, respectively, and ELISA also detected antibodies in chickens infected with A. paragallinarum with a sensitivity higher than that of HI tests. Taken together, the ELISA technique developed in this study is a valuable tool for the measurement of antibodies produced against the infectious coryza vaccine or in response to an infection with A. paragallinarum.

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A comparison of a blocking ELISA and a haemagglutination inhibition assay for the detection of antibodies to Avibacterium (Haemophilus) paragallinarum in sera from artificially infected chickens

Cited by 4 publications

References 11 publications

Development of Elisa Test Kit for Antibody Against Avibacterium Paragallinarum

Development of Elisa Test Kit for Antibody Against Avibacterium Paragallinarum

Haemophilus

Development of an Enzyme-Linked Immunosorbent Assay for the Measurement of Antibodies Against Infectious Coryza Vaccine

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