Silver enhanced nano-gold dot blot immunoassay for leptospirosis

Veerapandian, Raja; Prasad, Muthu; Bothammal, Palanisamy; Saranya, Perumal; Sumaiya, Krishnamoorthi; Mercy, Charles Solomon Akino; Natarajaseenivasan, Kalimuthusamy

doi:10.1016/j.mimet.2018.11.021

Cited by 6 publications

(3 citation statements)

References 10 publications

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“…These results are consistent with the previous studies of anti-rLipL32 antibodies produced using three peptide chains (EP3, EP4, and EP6), which showed specific immunoreactivity in the main band with a molecular size of 32-kDa in all Leptospira strains used [ 13 ]. Anti-rLipL32 antibody has also been used in a nanogold-based dot-blot immunoassay and proved to be a sensitive, rapid, and reliable method for early leptospirosis detection [ 41 ].…”

Section: Discussionmentioning

confidence: 99%

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Susanti,

Sudarmono,

Dharmayanti

et al. 2024

Vet World

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Background and Aim: Microscopic agglutination test (MAT) for the diagnosis of leptospirosis requires live cultures and is serovar-specific, while polymerase chain reaction (PCR) requires expensive equipment and sample preparation. The rLipL32 protein is conserved and can be used for the production of immunoglobulin G (IgG) anti-rLipL32 antibody, which can be used as a biomarker for leptospirosis diagnosis. This study aimed to produce and characterize an IgG anti-rLipL32 antibody as a biomarker for leptospirosis diagnosis. Materials and Methods: Escherichia coli rLipL32 was cultured and analyzed by PCR and sequencing. Cultures were used for rLipL32 protein expression and purification and the rLipL32 protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The rLipL32 protein was used to produce anti-rLipL32 serum and was analyzed by enzyme-linked immunosorbent assay (ELISA). Serum was purified to obtain IgG anti-rLipL32 antibody and characterized by SDS-PAGE and western blotting. Results: PCR was able to amplify the LipL32 gene from E. coli rLipL32, and sequencing analysis showed 99.19% similarity with pathogenic Leptospira. SDS-PAGE analysis showed a 32-kDa band. ELISA results showed an increase in OD in anti-rLipL32 serum compared to preimmune serum. Western blotting results showed that the IgG anti-rLipL32 antibody was able to bind and cross-reacts with pathogenic Leptospira serovar but not with E. coli or Staphylococcus aureus. Conclusion: IgG anti-rLipL32 antibody has high specificity and sensitivity against Leptospira pathogens. These findings suggest that IgG anti-rLipL32 antibody is a promising biomarker for the diagnosis of leptospirosis. Keywords: anti-rLipL32 serum, immunoglobulin G anti-rLipL32 antibody, Leptospira, rLipL32 protein.

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Section: Discussionmentioning

confidence: 99%

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Susanti,

Sudarmono,

Dharmayanti

et al. 2024

Vet World

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“…However, its applications are greatly restricted by the time-consuming and laborious steps. As a promising alternative, the dot-blot immunoassay has received considerable attention. − This method offers several significant advantages over traditional immunoassays, such as simple operation, low cost, and fast response time and is suitable for point-of-care tests. In addition, the small sample amount used in the assay renders this technique superior to other techniques for trace sample analysis.…”

Section: Introductionmentioning

confidence: 99%

Immunoglobulin G-Encapsulated Gold Nanoclusters as Fluorescent Tags for Dot-Blot Immunoassays

Zhuang

Deng

et al. 2019

ACS Appl. Mater. Interfaces

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Few-atom gold nanoclusters (AuNCs) have been fabricated and used for various fields owing to their remarkable optical and photophysical features. However, the rational design for the antibody-mediated synthesis of fluorescent AuNCs for direct antigen–antibody reactions remains unexplored. In this work, immunoglobulin G (IgG)-functionalized AuNCs (IgG-AuNCs) were successfully prepared via a facile and fast biomineralization process. The generated IgG-AuNCs can emit intense red fluorescence with a high photoluminescence quantum yield. Besides strong emission, the bioactivity of IgG on the IgG-AuNCs can be retained. Surface plasmon resonance measurements suggested that IgG-AuNCs can bind to goat anti-human IgG with an affinity constant of 6.21 × 10–8 M. A simple detection method was then developed using a dot-blot immunoassay with IgG-AuNCs as fluorescent tags. Experimental results confirmed that the IgG-AuNC-based fluorescent reporters had many advantages such as low nonspecific adsorption and good photostability, offering immense potential for the development of efficient biosensors. This work can be extended to other specific antibodies to produce multifunctional AuNCs and utilized to detect and monitor targeted analytes and biological events of interest.

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“…Dot blot hybridization (DBH) is a method of nucleic acid molecular hybridization, which has been widely used in virus detection in recent years. [31][32][33] DBH technology refers to the hybridization of nucleic acid probe with nucleic acid DNA of the sample to be tested on the hybridization membrane. The nucleic acid probe is equipped with appropriate markers for post-reaction detection, and combines with specific target molecules to form a hybrid body, which is then displayed by color reaction.…”

mentioning

confidence: 99%

A Dual-Amplification Electrochemical Aptasensor for Profenofos Detection

Zhang

Sun

Cheng

et al. 2020

J. Electrochem. Soc.

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The present study reported a dual-amplification electrochemical aptasensor for sensitive detection of profenofos (PFF) in vegetables. A screen-printed carbon electrode (SPCE) modified with graphitized multi-walled carbon nanotubes (MWCNTGr) and Au nanoshell was used as a test platform, which ensured a rapid detection process and showed a favorable electrochemical performance. MWCNTGr and Au nanoshell enhanced the electrical conductivity and the surface area, thus the detection signal was amplified. The affinity between PFF and its aptamer (Apt) was verified firstly by dot blot hybridization (DBH), and the result was exciting. Furthermore, the effects of the aptamers modified respectively with -NH2 and -SH on the current signal were compared with each other by cyclic voltammetry (CV), and results showed that the aptamers modified with -NH2 made the current signal change more obvious. Based on all above, a high-efficiency electrochemical aptasensor was fabricated with a wide linear range from 0.1–1 × 105 ng · ml−1 and a detection limit of 0.052 ng · ml−1 under the optimized conditions. This aptasensor had great specificity, stability and reproducibility. Hence, the developed aptasensor was successfully used to detect PFF in vegetables. The proposed method also has a potential for the detection of other organophosphorus pesticide (OPs).

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Silver enhanced nano-gold dot blot immunoassay for leptospirosis

Cited by 6 publications

References 10 publications

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Immunoglobulin G-Encapsulated Gold Nanoclusters as Fluorescent Tags for Dot-Blot Immunoassays

A Dual-Amplification Electrochemical Aptasensor for Profenofos Detection

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