Double‐Immunodiffusion Assay for Detecting Specific Antibodies (Ouchterlony)

Hornbeck, Peter

doi:10.1002/cpim.18

Cited by 18 publications

(16 citation statements)

References 3 publications

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“…The ODIT is the most useful serological technique for the detection of specific antibodies in sera, or for identifying viral antigen or antibodies in bursa. The ODIT is used to detect prominent white line of precipitation between bursal homogenates of the peripheral wells and known positive anti-IBDV hyper-immune serum of the central well due to antigen and antibody reaction (Hornbeck, 2017). Our data demonstrated that, in case of ODIT, hemorrhagic and edematous bursal homogenates were found positive for IBDV.…”

Section: Resultsmentioning

confidence: 67%

“…ODIT is also familiar as passive double immunodiffusion or AGID test. ODIT is a simple method to detect specific antibody and measure antigenicity (Hornbeck, 2017). It is simplest but least sensitive serological method (Sali, 2019).…”

Section: Introductionmentioning

confidence: 99%

“…RT-PCR is the method which helps for precise and early diagnosis of viruses from field samples (Hasan et al, 2010). Among these diagnostic methods, in this study we chose ODIT and RT-PCR as ODIT is simplest of all, inexpensive and has the ability for detecting specific antibodies (Dey et al, 2019;Hornbeck, 2017;Ouchterlony, 1948;Ouchterlony and Nilsson, 1986) and RT-PCR is specific, sensitive, speedy, reliable and accurate techniques for detection of IBD viruses (Hasan et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Efficacy of two diagnostic tests for the detection of infectious bursal disease viruses in chicken from different types of bursal samples

Afrin¹,

Chowdhury²,

Cho³

et al. 2020

Afr. J. Microbiol. Res.

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Infectious Bursal Disease (IBD) is an economically important irreversible immunosuppressive disease of young birds. The present study was designed to confirm the efficacy of two common diagnostic tests for the detection of Infectious Bursal Disease Virus (IBDV) from the three types of bursal samples collected from a recent outbreak in layer and broiler chickens of Gazipur district, Bangladesh. This study compared the degree of sensitivity between Ouchterlony Double Immunodiffusion Test (ODIT)and Reverse Transcription Polymerase Chain Reaction (RT-PCR) for the detection of IBD viral antigen from the bursal samples. A total of 180 field bursal samples (80 broiler and 100 from layer chicken) from suspected IBDV infected dead chickens were collected from 50 different poultry farms. Bursal homogenates were used to detect IBDV using ODIT and RT-PCR. Three types of bursal samples, hemorrhagic bursa (90), edematous bursa (78) and atrophied bursa (12) were selected for the detection of viral antigen. A panel of anti-sera and IBDV specific primer for VP2 gene was used for RT-PCR. The data demonstrated that, out of 180 field samples, 164 (91%) were positive by RT-PCR and 120 (67%) were positive by ODIT. Haemorrhagic bursas were more sensitive compared to oedematous bursas while no virus was detected from the samples of atrophied bursa. This study demonstrated that, RT-PCR was more sensitive and effective diagnostic tool compared to that of ODIT.

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Section: Resultsmentioning

confidence: 67%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Efficacy of two diagnostic tests for the detection of infectious bursal disease viruses in chicken from different types of bursal samples

Afrin¹,

Chowdhury²,

Cho³

et al. 2020

Afr. J. Microbiol. Res.

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“…Excellent performance in double immunodiffusion (DID) assay by using mini gel cassette DID was commonly used as a screening test for the monitoring of binding of antibody and antigen [53]. For conventional DID assay, the sample wells in gel are typically prepared by punching such as multiple-well patterns after the gel is solidified.…”

Section: Resultsmentioning

confidence: 99%

SELEX tool: a novel and convenient gel-based diffusion method for monitoring of aptamer-target binding

et al. 2020

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Background: Aptamers, single-stranded DNAs or RNAs, can be selected from a library containing random sequences using a method called Systematic Evolution of Ligands by EXponential Enrichment (SELEX). In SELEX, monitoring the enriching statuses of aptamer candidates during the process is a key step until today. Conformational change of an aptamer caused by target-binding in gel can be used to indicate its statuses of binding. Results: In this study, an easy-to-implement gel-based diffusion method (GBDM) was developed to monitor the interaction between enriched aptamer candidates and their targets. In order to prove the concept, characterization of aptamers targeting their targets including protein (thrombin) and non-protein molecules (acetamiprid, ATP, atrazine, profenofos and roxithromycin), respectively, were performed using mini gels. Our method has advantages over the common methods including easy performed with labor-and time-saving in experimental operation. The concept has been proven by monitoring enrichment of dynamic aptamer candidate libraries targeting a small molecule 2,2-bis(4-chlorophenyl) acetic acid (DDA) during SELEX process. A mini gel cassette was designed and fabricated by our laboratory to make mini agarose gels for diffusion with different directions. Conclusions:These results indicate that GBDM, in particular, chasing diffusion is suitable for monitoring the interaction between enriched aptamer candidates and their targets. These pioneering efforts are helpful for novel aptamer selection by breaking through the technical bottleneck of aptamer development and helpful for development of novel aptasensors.

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“…The method used here was first described by Ouchterlony in 1948 42 . It is a classic and simple technique that evaluates antibodies in animal or human sera 43,44 .…”

Section: Methodsmentioning

confidence: 99%

Chemosynthesis and characterization of site-specific N-terminally PEGylated Alpha-momorcharin as apotential agent

Sun

Zhang

et al. 2018

Sci Rep

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Alpha-momorcharin (α-MC), a type I ribosome-inactivating protein (RIP) isolated from Momordica charantia seeds, has been extensively studied for its antitumor, antiviral and antifungal activities. However, as an exogenous protein, problems associated with short half-life and strong immunogenicity have limited its clinical application. Poly (ethylene glycol) (PEG), as a polyether compound, is a well established and efficient modifier to develop it as a potential agent. Nevertheless, conventional PEGylation is not site-controlled and the conjugates are often not homogenous due to the generation of multi-PEGylated derivatives. To obtain a homogenous mono-PEGylated α-MC, the PEGylation was carried out by coupling a 20 kDa mPEG-butyraldehyde (mPEG-ALD) with α-MC. The product was separated and purified by MacroCap SP chromatography. Results from SDS-PAGE and MALDI-TOF MS revealed that the PEGylated α-MC consisted of one molecule mPEG and α-MC. Edman degradation confirmed that the N-terminal residue of α-MC was successfully coupled with mPEG-ALD. The mono-PEGylated α-MC possessed an extremely similar secondary structure to native α-MC through spectral analyses. In addition, it also showed low immunogenicity by double immunodiffusion and preserved moderate antitumor activity to three kinds of tumor cell lines in vitro. Finally, trypsin resistance was also considerably improved.

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Double‐Immunodiffusion Assay for Detecting Specific Antibodies (Ouchterlony)

Cited by 18 publications

References 3 publications

Efficacy of two diagnostic tests for the detection of infectious bursal disease viruses in chicken from different types of bursal samples

Efficacy of two diagnostic tests for the detection of infectious bursal disease viruses in chicken from different types of bursal samples

SELEX tool: a novel and convenient gel-based diffusion method for monitoring of aptamer-target binding

Chemosynthesis and characterization of site-specific N-terminally PEGylated Alpha-momorcharin as apotential agent

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