Emzyme-Linked Immunosorbent Assay for Detection of Canine Chromogranin A by Use of Immunological Cross-Reactivity of Rabbit Anti-Bovine Chromogranin A Antibody

Akiyoshi, Hideo; Aoki, Mica; Kumagai, D.; Saleh, Nahed; Noda, Katsura; Shimada, Terumasa; Sugii, Shunji; Ohashi, Fumihito

doi:10.1292/jvms.67.535

Cited by 5 publications

(3 citation statements)

References 17 publications

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“…The precision obtained in our assay was higher to those established for ELISA techniques for CgA measure in human saliva (,9% for intra-assay CV and ,13% for inter-assay CV; Filaire et al, 2009) and much better than those reported for canine CgA measurements by ELISA (,16% for intra-assay CV and ,17% for inter-assay CV; Akiyoshi et al, 2005b). Such a low intra-assay variation makes assays of duplicate samples unnecessary, yielding savings in time and sample volume required to perform the assay.…”

Section: Discussioncontrasting

confidence: 56%

Measurement of chromogranin A in porcine saliva: validation of a time-resolved immunofluorometric assay and evaluation of its application as a marker of acute stress

Escribano

Soler

Gutiérrez

et al. 2013

Animal

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The objective of this study was to develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for porcine salivary chromogranin A (CgA) measurements, using a species-specific antibody, and evaluate its behaviour in an acute stress model. Polyclonal antibodies were produced in rabbits immunized with a synthetic porcine fragment of CgA 3592379 and used to develop a sandwich TR-IFMA. This TR-IFMA was analytically validated and showed intra-and inter-assay coefficients of variation of 6.23% and 5.82%, respectively, an analytical limit of detection of 4.27 3 10 23 mg/ml and a limit of quantification of 24.5 3 10 23 mg/ml. The assay also demonstrated a high level of accuracy, as determined by linearity under dilution (r 5 0.975) and recovery tests. When a model of experimental acute stress, in which animals were immobilized for 3 min with a nose snare (stressor stimulus), was applied, a significant increase (P , 0.05) in CgA levels in saliva was detected at 15 min post-stressor stimulus. These results indicate that the assay developed in this study could measure CgA in porcine saliva in a reliable way and that the concentrations of CgA in saliva samples of pigs increase after an acute stress situation.

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Section: Discussioncontrasting

confidence: 56%

Measurement of chromogranin A in porcine saliva: validation of a time-resolved immunofluorometric assay and evaluation of its application as a marker of acute stress

Escribano

Soler

Gutiérrez

et al. 2013

Animal

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“…Within the tested incubation periods, the longer incubation period made the displacement curve steep and left-shifted, suggesting both better precision and higher sensitivity (Figure 2a,b). The assay sensitivity, calculated as 0.46 nM, seems to be sufficiently lower than the plasma concentrations of CgA in other species (Akiyoshi et al, 2005;Myers et al, 1997;Schneider et al, 2018). Attempts to duplicate the dosedependent decrease of optical density with serially diluted recombinant bCgA revealed an apparent parallelism to the peptide-based displacement curve.…”

Section: Discussionmentioning

confidence: 99%

Development of enzyme immunoassay for chromogranin A (CgA) and profiling of plasma CgA concentrations in the cow

Saito

Ido-Matsumoto

Kumazaki

et al. 2021

Animal Science Journal

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The objectives of this study were to establish and characterize a homologous immunoassay for bovine chromogranin A (bCgA) and to profile plasma bCgA concentrations during early pregnancies. We synthesized oligopeptide corresponding to the amino acid sequence 341–355 of bCgA for immunizing rabbits and peptide corresponding to the amino acid sequence 336–365 of bCgA for both a biotinylated tracer and reference standards. Recombinant bCgA protein was also generated in Escherichia coli lysate. Dose‐dependent displacement curves were obtained from 1 to 1,000 nM of the reference standards. The displacement curves showed a good relationship between the reference standards of the synthetic peptide and the serially diluted plasma sample or recombinant bCgA protein generated in the present study. The assay sensitivity defined as the value of two standard deviations below the zero standard was calculated as 0.46 nM. The intraassay and interassay coefficients of variation were 6.48% and 13.4%, respectively. Changes in the plasma bCgA concentrations in early pregnancies undulated in nonpregnant animals. The results of the present study suggest that assaying plasma bCgA concentrations could be utilized as measures to evaluate the physiological status of cattle.

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“…Chromogranin A (CgA) in peripheral samples is a known SAM activation marker in humans, dogs and pigs [11][12][13][14]. CgA is an acidic protein that is stored in the secretory granules of endocrine, neuroendocrine and neuronal cells [15,16].…”

Section: Introductionmentioning

confidence: 99%

Salivary Chromogranin A (CgA) Response to the Noradrenaline Transporter Blocker Atomoxetine in Dogs

Kooriyama

Mukhopadhyay

Moore

et al. 2021

Animals

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Since salivary chromogranin A (CgA) is one of the known sympathetic adrenomedullar system (SAM) stress markers in humans and pigs, this study aimed to investigate whether salivary CgA in dogs reflects SAM activation. Our hypothesis was that salivary CgA would increase when central noradrenaline was pharmacologically induced. A selective noradrenaline transporter blocker, atomoxetine, was orally administered without causing any aversive responses in nine laboratory dogs to see if it would increase salivary CgA. Three treatment groups (i.e., atomoxetine, placebo, and pre-administration of a selective alpha-2 adrenoreceptor agonist (dexmedetomidine) followed by atomoxetine) were prepared with a randomized crossover design. Saliva sample collection, heart rate measurement and behavior observation were performed at Time 0 (baseline) and at 30, 60, 90 and 150 min after each treatment administration. The results demonstrated that salivary CgA significantly increased at 90 and 150 min in the atomoxetine treatment (p < 0.05), whereas it was not observed in the other two treatments. The present study showed that salivary CgA was increased by atomoxetine-induced SAM activation. However, this increase was blocked if dexmedetomidine was pre-administered. Overall, the results indicate that salivary CgA is a potential candidate for SAM-mediated stress markers in dogs. Further study to determine the dynamics of salivary CgA will be helpful in its practical use.

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Emzyme-Linked Immunosorbent Assay for Detection of Canine Chromogranin A by Use of Immunological Cross-Reactivity of Rabbit Anti-Bovine Chromogranin A Antibody

Cited by 5 publications

References 17 publications

Measurement of chromogranin A in porcine saliva: validation of a time-resolved immunofluorometric assay and evaluation of its application as a marker of acute stress

Measurement of chromogranin A in porcine saliva: validation of a time-resolved immunofluorometric assay and evaluation of its application as a marker of acute stress

Development of enzyme immunoassay for chromogranin A (CgA) and profiling of plasma CgA concentrations in the cow

Salivary Chromogranin A (CgA) Response to the Noradrenaline Transporter Blocker Atomoxetine in Dogs

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