Development of monoclonal antibody based sandwich ELISA for the rapid detection of pathogenic Vibrio parahaemolyticus in seafood

Kumar, Ballamoole Krishna; Raghunath, Pendru; Devegowda, Devananda; Deekshit, Vijaya Kumar; Venugopal, M. N.; Karunasagar, Iddya

doi:10.1016/j.ijfoodmicro.2010.12.030

Cited by 95 publications

(45 citation statements)

References 29 publications

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“…positive (Johnson, 2009). Another virulence factor, the tdh-related hemolysin (trh) is generally associated with the Kanagawa phenomenon negative (KP -) strains or with urease positive strains of V. parahaemolyticus (Okuda et al, 1997;Kumar et al, 2011). Though most of the environmental strains of V. parahaemolyticus are typically not human pathogens, they cause disease in fish and shellfish Nordstrom and DePaola., 2003;MartinezUrtaza et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

“…ELISA techniques are widely used immunodiagnostic techniques to detect pathogens in culture systems (Romestand et al, 1993;Kumar et al, 2011) due to its sensitivity. In the latex agglutination test for the rapid identification of V. parahaemolyticus (Chang et al, 1994), two bacterial outer membrane proteins, with molecular weights of 36 kDa and 34 kDa were used as antigens for producing polyclonal antibodies.…”

Section: Discussionmentioning

confidence: 99%

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Selection of specific cell wall antigen for rapid detection of fish pathogenic Vibrio parahaemolyticus by enzyme immunoassay

Anand

Sobhana²,

George³

et al. 2016

Indian J. Fish.

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An enzyme linked immunosorbant assay (ELISA) was developed, using polyclonal antibodies against a specific cell surface protein of Vibrio parahaemolyticus, for rapid detection of the organism. Nine virulent strains and one type strain of V. parahaemolyticus, one strain each of Vibrio vulnificus and Vibrio alginolyticus were used for the study. Cell surface proteins were extracted from all the strains and were analysed by SDS PAGE. One distinct band with molecular weight of 34 kDa, abundant in all the V. parahaemolyticus strains and lacks in other Vibrio species, was selected as cell wall antigen for immunisation. Polyclonal antibodies were raised against the selected 34 kDa protein of V. parahaemolyticus after preparative electrophoresis. An indirect plate ELISA was developed using this antiserum for detection of crude cell surface protein as well as whole cells of V. parahaemolyticus. All the 9 virulent strains and one type strain of V. parahaemolyticus tested, produced positive results, using the ELISA technique. To assess the specificity of the polyclonal serum, cross reaction studies with other Vibrio species such as V. vulnificus and V. alginolyticus were conducted by indirect plate ELISA. The results have clearly shown that antibodies directed against 34 kDa cell surface protein can be used for specific detection of fish pathogenic V. parahaemolyticus.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Selection of specific cell wall antigen for rapid detection of fish pathogenic Vibrio parahaemolyticus by enzyme immunoassay

Anand

Sobhana²,

George³

et al. 2016

Indian J. Fish.

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“…[5][6][7] Although conventional culturing ensures the accuracy of determinations, and are optimum methods for VP identification, there still exist some shortcomings, such as long-time consumption and cumbersome practical application. Various approaches have been developed to obtain better performances of VP analysis, including enzyme-linked immunesorbent assay (ELISA), [8][9][10] DNA probe, 11 most probable number (MPN), 12,13 polymerase chain reaction (PCR), [14][15][16][17][18] loop-mediated isothermal amplification (LAMP), [19][20][21][22][23] and electrochemistry (EC). 24,25 Despite each of these new approaches having a combination of excellent sensitivity, accuracy and specificity, they still suffer drawbacks involving high analytical cost, need for expensive equipment, and professional trained personnel.…”

Section: Introductionmentioning

confidence: 99%

Christmas-tree Derived Amplification Immuno-Strategy for Sensitive Visual Detection of Vibrio parahaemolyticus Based on Gold Label Silver Stain Technology

Song

et al. 2017

ANAL. SCI.

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Recently, Liu and his coworkers designed a Fe(III)-grafted g-C3N4 photocatalyst that displayed an excellent photocatalytic performance. 37 These make it promising in the development of biosensors with high 2017 © The Japan Society for Analytical Chemistry † To whom correspondence should be addressed. A developed Christmas-tree derived immunosensor based on a gold label silver stain (GLSS) technique was fabricated for a highly sensitive analysis of Vibrio parahaemolyticu (VP). In this strategy, captured VP antibody (cAb) was immobilized on a solid substrate; then, the VPs were sequentially tagged with a signal probe by incubating the assay with a detection VP antibody (dAb) conjugated gold nanoparticles (AuNPs)-labeled graphite-like carbon nitride (g-C3N4). Finally, the attached signal probe could harvest a visible signal by the silver meal deposition, and then followed by homebrew Matlab 6.0 as a grey value acquisition. In addition, the overall design of the biosensor was established in abundant AuNPs and g-C3N4 with a two-dimensional structure, affording a bulb-decorated Christmas-tree model. Moreover, with the optimized conditions, the detection limit of the as-proposed biosensor is as low as 10 2 CFU (Colony-Forming Units) mL -1 , exhibiting an increase of two orders of magnitude compared with the traditional immune-gold method. Additionally, the developed visible immunosensor was also successfully applied to the analysis of complicated samples.

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“…To improve efficiency, more rapid assays were developed for detection of these pathogens in oysters using molecular approaches [7,[14][15][16] and immunoassays [17,18]. The molecular assays such as PCR have been used widely because of their high sensitivity, speed, and convenience [7,19,20]. Efficient amounts of sample DNA of high quality is of critical importance for molecular assays.…”

Section: Introductionmentioning

confidence: 99%

“…Efficient amounts of sample DNA of high quality is of critical importance for molecular assays. So far, different methods have been used to extract the DNA samples, including commercial kits [19,21,22], the phenol-chloroform method [15,23] and the boiling lysis method [10,24,25].…”

Section: Introductionmentioning

confidence: 99%

http://www.omicsonline.org/open-access/overcoming-pseudomonas-aeruginosa-resistance-caused-by-glycocalyx-with-tobracef-1948-5948.1000145.php?aid=25925

Dai¹

2014

J Microb Biochem Technol

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Oysters are filter feeders that bioaccumulate bacteria in water while feeding. To evaluate the bacterial genomic DNA extracted from retail oyster tissues, including the gills and digestive glands, four isolation methods were used. Genomic DNA extraction was performed using the Allmag™ Blood Genomic DNA (Allrun, Shanghai, China), the MiniBEST Bacterial Genomic DNA Extraction kits (Takara, Dalian, China), and the phenol-chloroform and boiling lysis methods. The concentration of the genomic DNA was measured using a spectrophotometer. The purity of the genomic DNA was evaluated by PCR amplification of 16S rDNA followed by determining the cloning efficiency of the amplicon into the pMD19-T vector. Furthermore, the bacterial DNA quality was also evaluated by PCR assays using a pair of species-specific primers for Vibrio parahaemolyticus. Our results showed that the two commercial kits produced the highest purity of DNA, but with the lowest yields. The phenol-chloroform method produced the highest yield although it was time-consuming. The boiling lysis method was simple and cost effective; however, it was only suitable to isolate genomic DNA from bacteria present in retail samples following an enrichment step. The two commercial kits were good candidates for genomic DNA extraction from retail oyster tissues without enrichment.

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Development of monoclonal antibody based sandwich ELISA for the rapid detection of pathogenic Vibrio parahaemolyticus in seafood

Cited by 95 publications

References 29 publications

Selection of specific cell wall antigen for rapid detection of fish pathogenic Vibrio parahaemolyticus by enzyme immunoassay

Selection of specific cell wall antigen for rapid detection of fish pathogenic Vibrio parahaemolyticus by enzyme immunoassay

Christmas-tree Derived Amplification Immuno-Strategy for Sensitive Visual Detection of Vibrio parahaemolyticus Based on Gold Label Silver Stain Technology

http://www.omicsonline.org/open-access/overcoming-pseudomonas-aeruginosa-resistance-caused-by-glycocalyx-with-tobracef-1948-5948.1000145.php?aid=25925

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