Elimination of shrimp endogenous alkaline phosphatase background and development of enzyme immunoassays for the detection of white spot syndrome virus (WSSV)

Zhan, Wenbin; Wang, Xiaojie; Chen, Jing; Xing, Jing; Fukuda, Hideo

doi:10.1016/j.aquaculture.2004.05.042

Cited by 25 publications

(14 citation statements)

References 7 publications

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“…A c c e p t e d M a n u s c r i p t 12 Phosphatase hydrolysis of p-NPP by alkaline phosphatase catalysis follows the Michaelis-Menten equation. As reported in Table 2, the values of k cat , K m , and k cat /K m measured from Lineweaver Burk plot, were 95 s -1 , 0.3 µM and 316.6 s -1 µM -1 , respectively.…”

Section: Extraction and Purification Of The Enzymementioning

confidence: 99%

“…A relatively low thermostability can be useful for AP removing from the reaction mixture without additional purification in DNA techniques [12]. Furthermore, Fenneropenaeus merguiensis alkaline phosphatase can be a potential marker for specific plasma membrane purification, additionaly this enzyme can be used as a marker to differentiate testicular origin azoospermia or oligospermia from ejaculatory failure, as well as ejaculatory failure and excurrent duct blockages in an inexpensive way [8][9][10][11][12].…”

Section: Extraction and Purification Of The Enzymementioning

confidence: 99%

“…Also their application for the determination of zinc in insulin to control injection quality and magnesium in human urine for the diagnosis and treatment of magnesium deficiency was shown [11]. Marine organism have been shown to be a valuable source for the isolation of novel APs with potential practical application [12]. Current report has novelty as it explained for the first time kinetics and thermodynamics of soluble p-NPP hydrolysis and irreversible inactivation of alkaline phosphatase from Fenneropenaeus merguiensis brain and compatibility with other enzymes to exploit its efficiency in a variety of medicine and biotechnology applications.…”

Section: Introductionmentioning

confidence: 98%

“…This enzyme is routinely used as a labeling enzyme in electrochemical enzyme immunoassays and immunosensors and numerous approaches have been reported for determining herbicides, pesticides, and different biologically active organic compounds [10][11][12]. Additionally, prospects of using different alkaline phosphatases bearing zinc and magnesium ions in their catalytic and allosteric sites, respectively, in pharmaceutical and clinical analysis were demonstrated.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Purification and biochemical properties of highly efficient alkaline phosphatase from Fenneropenaeus merguiensis brain

Homaei

2015

Journal of Molecular Catalysis B: Enzymatic

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Section: Extraction and Purification Of The Enzymementioning

confidence: 99%

Section: Extraction and Purification Of The Enzymementioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Purification and biochemical properties of highly efficient alkaline phosphatase from Fenneropenaeus merguiensis brain

Homaei

2015

Journal of Molecular Catalysis B: Enzymatic

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“…For example, shrimp endogenous peroxidase and AP were found to produce falsepositive results when ELISA and immunodot blot assay were used to detect the WSSV in shrimp tissue by using antibodies conjugated with AP and HRP [15,16]. In such cases, GOD would be a good candidate for a labeling enzyme.…”

Section: Introductionmentioning

confidence: 99%

Application of methyl parathion hydrolase (MPH) as a labeling enzyme

et al. 2008

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Methyl parathion hydrolase (MPH) is an enzyme that catalyzes the degradation of methyl parathion, generating a yellow product with specific absorption at 405 nm. The application of MPH as a new labeling enzyme was illustrated in this study. The key advantages of using MPH as a labeling enzyme are as follows: (1) unlike alkaline phosphatase (AP), horseradish peroxidase (HRP), and glucose oxidase (GOD), MPH is rarely found in animal cells, and it therefore produces less background noise; (2) its active form in solution is the monomer, with a molecular weight of 37 kDa; (3) its turnover number is 114.70±13.19 s −1 , which is sufficiently high to yield a significant signal for sensitive detection; and (4) its 3D structure is known and its C-terminal that is exposed to the surface can be easily subjected to the construction of genetic engineering monocloning antibody-enzyme fusion for enzyme-linked immunosorbent assay (ELISA). To demonstrate its utility, MPH was ligated to an single-chain variable fragment (scFv), known as A1E, against a white spot syndrome virus (WSSV) with the insertion of a [-(Gly-Ser) 5 -] linker peptide. The resulting fusion protein MPH-A1E possessed both the binding specificity of the scFv segment and the catalytic activity of the MPH segment. When MPH-A1E was used as an ELISA reagent, 25 ng purified WSSV was detected; this was similar to the detection sensitivity obtained using A1E scFv and the HRP/Anti-E Tag Conjugate protocol. The fusion protein also recognized the WSSV in 1 μL hemolymph from an infected shrimp and differentiated it from a healthy shrimp.

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Semiquantitative dot‐blot immunogold assay for specific detection of white spot syndrome virus

Bunsanong

Chotigeat

Deachamag

et al. 2018

Biotech and App Biochem

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A dot-blot immunogold assay (DBIA) was developed to detect white spot syndrome virus (WSSV) using the polyclonal antibody VP26 (anti-VP26). The anti-VP26 was immobilized on gold nanoparticles (Ab-AuNPs), and a nitrocellulose membrane was used as a detection pad. When the target WSSV bound to the Ab-AuNPs a reddish dot appeared on the surface of the membrane used within 2-5 Min, which could be seen with the naked eye. The test was able to detect WSSV at concentrations as low as 10 copies μL of WSSV. The DBIA developed had good specificity, and the colloidal gold probe can be applied within 2-3 days when stored at 4 °C. For real sample analysis, the DBIA was applied to samples of seawater used for shrimp cultivation without sample preparation. The results indicate that sample 1 showed a positive result, whereas samples 2 and 3 produced negative results. Then, samples 2 and 3 were spiked with WSSV for method validation. To confirm the performance of the DBIA developed, polymerase chain reaction (PCR) was conducted and the PCR results were the same as those found by the DBIA. Therefore, the DBIA developed could be applied for WSSV detection in real water samples.

show abstract

Elimination of shrimp endogenous alkaline phosphatase background and development of enzyme immunoassays for the detection of white spot syndrome virus (WSSV)

Cited by 25 publications

References 7 publications

Purification and biochemical properties of highly efficient alkaline phosphatase from Fenneropenaeus merguiensis brain

Purification and biochemical properties of highly efficient alkaline phosphatase from Fenneropenaeus merguiensis brain

Application of methyl parathion hydrolase (MPH) as a labeling enzyme

Semiquantitative dot‐blot immunogold assay for specific detection of white spot syndrome virus

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