Sensitivity improvement of rapid<i>Vibrio harveyi</i>detection with an enhanced chemiluminescent-based dot blot

Li, Hui; Xiao, Jingfan; Zhou, Ya; Wang, Qiyao; Zhang, Yuanxing

doi:10.1111/lam.12763

Cited by 5 publications

(2 citation statements)

References 18 publications

(27 reference statements)

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“…Serological approaches have garnered support. For example, a chemiluminescent-based dot blot incorporating monoclonal antibodies was described, and invoked a brief 2-h incubation leading to a limit of detection of 2 × 10 5 /ml; this was regarded as 50 times more sensitive than the conventional dot blot, and was superior (~ 1000 times more sensitive) to the indirect enzyme-linked immunosorbent assay (Li et al 2017).…”

Section: Diagnosismentioning

confidence: 99%

Vibrio harveyi: a serious pathogen of fish and invertebrates in mariculture

Zhang

Austin

2020

Mar Life Sci Technol

187

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Vibrio harveyi, which belongs to family Vibrionaceae of class Gammaproteobacteria, includes the species V. carchariae and V. trachuri as its junior synonyms. The organism is a well-recognized and serious bacterial pathogen of marine fish and invertebrates, including penaeid shrimp, in aquaculture. Diseased fish may exhibit a range of lesions, including eye lesions/blindness, gastro-enteritis, muscle necrosis, skin ulcers, and tail rot disease. In shrimp, V. harveyi is regarded as the etiological agent of luminous vibriosis in which affected animals glow in the dark. There is a second condition of shrimp known as Bolitas negricans where the digestive tract is filled with spheres of sloughed-off tissue. It is recognized that the pathogenicity mechanisms of V. harveyi may be different in fish and penaeid shrimp. In shrimp, the pathogenicity mechanisms involved the endotoxin lipopolysaccharide, and extracellular proteases, and interaction with bacteriophages. In fish, the pathogenicity mechanisms involved extracellular hemolysin (encoded by duplicate hemolysin genes), which was identified as a phospholipase B and could inactivate fish cells by apoptosis, via the caspase activation pathway. V. harveyi may enter the so-called viable but nonculturable (VBNC) state, and resuscitation of the VBNC cells may be an important reason for vibriosis outbreaks in aquaculture. Disease control measures center on dietary supplements (including probiotics), nonspecific immunostimulants, and vaccines and to a lesser extent antibiotics and other antimicrobial compounds.

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Section: Diagnosismentioning

confidence: 99%

Vibrio harveyi: a serious pathogen of fish and invertebrates in mariculture

Zhang

Austin

2020

Mar Life Sci Technol

187

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“…The specificity of the grouper IgM antibody was tested using the dot blot method with V. alginolyticus and V. harveyi antigens and the specific receptor organ of the fish [ 33 ]. First, nitrocellulose (NC) was soaked in PBS for 30 min in the dot blotting chamber.…”

Section: Methodsmentioning

confidence: 99%

In vivo test of Vibrio alginolyticus and Vibrio harveyi infection in the humpback grouper (Cromileptes altivelis) from East Java Indonesia

et al. 2022

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Background and Aim: The need for fish seeds resistant to bacterial and viral infections has encouraged studies on the molecular pathogenesis mechanism of Vibrio bacteria, such as Vibrio alginolyticus and Vibrio harveyi, regarding the receptor organs, protein adhesion mechanisms, and antibody responses of the humpback grouper. This study aims to confirm the characteristics of the specific proteins expressed in the receptor organ of the humpback grouper (Cromileptes altivelis) using the expression of V. alginolyticus and V. harveyi bacteria. Materials and Methods: The study was conducted by isolating crude protein and whole cells from both the Vibrio bacteria. In addition, serum and organ tissue were also isolated from fish samples. Then, hemagglutination and dot blot tests with polyacrylamide gel electrophoresis analysis were performed to determine the highest expression of receptor from the whole bacterial cells and crude protein from both healthy and infected (V. alginolyticus and V. harveyi) fishes. Scanning electron microscope results showed that V. alginolyticus and V. harveyi could express bundle-forming pili, which is involved in bacterial autoaggregation and the mediation of the initial attachment of bacteria to their host cells. Results: These results indicated that all the specific receptors for protein in fish organs recognized vibriosis antigens. The specificity test showed that the brain, eye, and kidney organs' receptors provided a quality and quantity level of responses at 22.63, 53.95, and 43.15 kDa, respectively. The polyclonal anti-V. alginolyticus immunoglobulin M (IgM) antibodies were more cross-reactive than the anti-V. harveyi IgM. Hence, this shows that V. alginolyticus bacteria are more pathogenic than V. harveyi. Conclusion: In the future, the molecular characteristics of V. alginolyticus and V. harveyi antigens and the specific receptor organ proteins in the humpback grouper can be developed as the basis for constructing molecular peptide-based vaccine materials.

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