Detection and estimation of avian infectious bronchitis virus antigen by a novel indirect liquid-phase blocking enzyme-linked immunosorbent assay using chicken and rabbit affinity purified immunoglobulins

Lougovskaia, Natalia N; Lougovskoi, Andrei A; Bochkov, Yury A.; Batchenko, Galina V; Мудрак, Н. С.; Drygin, V. V.; Борисов, А. В.; Borisov, Vladimir; Gusev, A. A.

doi:10.1080/0307945021000024571

Cited by 9 publications

(8 citation statements)

References 27 publications

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“…Total RNA was isolated from 50 ll of the sample using GF/F glass fiber filters as described previously [21,22]. Briefly, the extraction procedure was as follows: 0.5 ml of 4 M guanidine isothiocyanate (Fluka) was added to 50 ll of the sample; the mixture was vortexed and incubated at room temperature for 15-20 min.…”

Section: Rna Isolationmentioning

confidence: 99%

Phylogenetic analysis of partial S1 and N gene sequences of infectious bronchitis virus isolates from Italy revealed genetic diversity and recombination

et al. 2006

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A total of ten infectious bronchitis virus (IBV) isolates collected from commercial chickens in Italy in 1999 were characterized by RT-PCR and sequencing of the S1 and N genes. Phylogenetic analysis based on partial S1 gene sequences showed that five field viruses clustered together with 793/B-type strains, having 91.3-98.5% nucleotide identity within the group, and one isolate had very close sequence relationship (94.6% identity) with 624/I strain. These two IBV types have been identified in Italy previously. The other three variant isolates formed novel genotype detected recently in many countries of Western Europe. For one of these variant viruses, Italy-02, which afterwards became the prototype strain, the entire S1 gene was sequenced to confirm its originality. In contrast, phylogenetic analysis of more conserved partial N gene sequences, comprising 1-300 nucleotides, revealed different clustering. Thus, three variant IBVs of novel Italy-02 genotype, which had 96.7-99.2% S1 gene nucleotide identity with each other, belonged to three separate subgroups based on N gene sequences. 624/I-type isolate Italy-06 together with Italy-03, which was undetectable using S1 gene primers, shared 97.7% and 99.3% identity, respectively, in N gene region with vaccine strain H120. Only one of the 793/B-type isolates, Italy-10, clustered with the 793/B strain sharing 99.3% partial N gene identity, whereas the other four isolates were genetically distant from them (only 87.7-89.7% identity) and formed separate homogenous subgroup. The results demonstrated that both mutations and recombination events could contribute to the genetic diversity of the Italian isolates.

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Section: Rna Isolationmentioning

confidence: 99%

Phylogenetic analysis of partial S1 and N gene sequences of infectious bronchitis virus isolates from Italy revealed genetic diversity and recombination

et al. 2006

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“…The detection limit of IBV in the Con A-S-ELISA (10 5.5 EID 50 /mL) was similar to the levels reported previously for conventional S-ELISA and competitive ELISA (1,15,19,34). However, this level is higher than the detection limit found, in monoclonal antibody-S-ELISA (12), probably because the characteristics of anti-IBV monoclonal antibodies (mAbs) used as capture reagent by these authors.…”

Section: Discussionmentioning

confidence: 45%

“…Enzyme Linked Immunosorbent Assays (ELISAs) have been developed and proved their efficacy, either for the detection of IBV antigen (1,12,15,19,21,34), or anti-IBV antibodies (2,3,4,8,16,22,29).…”

Section: Introduction I Nfectious Bronchitis Virus (Ibv) Is a Member Ofmentioning

confidence: 99%

Detection of Infectious Bronchitis Virus and Specific Anti- Viral Antibodies Using a Concanavalin A–Sandwich–ELISA

Bronzoni

Fátima²,

Montassier

et al. 2005

Viral Immunology

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Concanavalin A-Sandwich ELISA (Con A-S-ELISA) was developed for the detection of infectious bronchitis virus (IBV) or chicken specific anti-viral antibodies. The antigen detection limit for the Con A-S-ELISA was 10 5,1 EID 50 /mL. Three homologous and four heterologous IBV strains were similarly detected. This assay was highly effective in detecting the virus after infected tissue homogenates were passed once in embryonated chicken eggs, showing a good agreement with virus isolation technique. The Con A-S-ELISA was also used to measure anti-IBV chicken antibodies and showed a high coefficient of correlation (r ‫؍‬ 0.85) and an agreement of k ‫؍‬ 0.80 with the commercially available Indirect-ELISA. The relative sensitivity and specificity between these two tests were, respectively, 92.86% and 95.65% with an accuracy of 93.39%. Thus, the Con A-S-ELISA proved to be able to detect alternatively homologous and heterologous IBV strains or specific chicken anti-IBV antibodies, using the Con A as capture reagent of this assay. 569

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“…Traditional methods of measuring PFU or TCID 50 (EID 50 ) are subjective, tedious and time-consuming, which leads to variabilities in the quality and timely production of the vaccine. Although other titration methods have been explored to solve these problems, such as flow cytometry, enzyme-linked immunosorbent assay and fluorescence detection of virus-induced apoptosis, 7,18,19 they also have disadvantages such as being poorly sensitivity or not suitable for trivalent LAIVs. The RT-qPCR method has gained increased attention and is expected to overcome drawbacks of the conventional assays.…”

Section: Discussionmentioning

confidence: 99%

Development of one-step real-time PCR assay for titrating trivalent live attenuated influenza vaccines

Zang

Ge³

et al. 2014

Human Vaccines & Immunotherapeutics

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Traditionally, infectivity of a trivalent live attenuated influenza vaccines (LAIVs) is titrated by determining the 50% egg infectious dose assay (EID50) or plaque forming units (PFU), which requires specific monoclonal antibodies to neutralize 2 strains while estimating the titer of the non-neutralized strain. Compared to this time-consuming, laborious, subjective and variable process, reverse transcription-quantitative real-time PCR (RT-qPCR) technology has advantages of rapidity, sensitivity, reproducibility and reduced contamination, thus has been applied widely for detecting pathogens and measuring viral titers. In this study, the critical harvest time was determined to be 18 h post-infection (hpi) for type A influenza and 12 hpi for type B influenza, but no significant difference between titers at 12 hpi and 18 hpi for the type B strain was observed. In conclusion, trivalent LAIVs can be titrated simultaneously within 24 h by this one-step RT-qPCR assay, which yielded titers comparable to those obtained by the traditional EID50 assay. Therefore, the RT-qPCR assay may be used as a highly specific, sensitive, precise and rapid alternative to the EID50 assay for titering LAIVs.

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Detection and estimation of avian infectious bronchitis virus antigen by a novel indirect liquid-phase blocking enzyme-linked immunosorbent assay using chicken and rabbit affinity purified immunoglobulins

Cited by 9 publications

References 27 publications

Phylogenetic analysis of partial S1 and N gene sequences of infectious bronchitis virus isolates from Italy revealed genetic diversity and recombination

Phylogenetic analysis of partial S1 and N gene sequences of infectious bronchitis virus isolates from Italy revealed genetic diversity and recombination

Detection of Infectious Bronchitis Virus and Specific Anti- Viral Antibodies Using a Concanavalin A–Sandwich–ELISA

Development of one-step real-time PCR assay for titrating trivalent live attenuated influenza vaccines

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