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A highly sensitive detection for foot-and-mouth disease virus by gold nanopariticle improved immuno-PCR
Yao-zhong Ding1, Yong-sheng Liu, Jian-hua Zhou
1Chinese Academy of Agricultural Sciences, Lanzhou, Gansu,China.
Background:
Foot-and-mouth disease (FMD) is one of the most contagious of all artiodactyl animal diseases, and its infection has an obvious ability to spread over long distances and to contribute to epidemics in FMD-free areas. A highly sensitive and specific method is required to detect FMDV. In this study, we evaluated the usefulness of a bio-barcode assay (BCA) technique for detecting clinical samples of FMDV.
Methods:
Highly sensitive gold nanopariticle (GNP) improved immuno -PCR (GNP-IPCR) which derived from the bio-barcode assay (BCA) was designed for the detection of FMDV. The target viral particles were captured by a polyclonal antibody coated on ELISA microplate, followed by adding GNP which was dually modified with oligonucleotides and a FMDV specific monoclonal antibody (MAb) 1D11 to form a sandwiched immune complex. After the formation of immuno-complex, the signal DNA was released by heating, and consequently characterized by PCR and real time PCR.
Results:
The detection limit of GNP-PCR could reach to 10 fg/ml purified FMDV particles, and the assay can detect clinical samples of FMDV with highly sensitivity, while detect limit of conventional ELISA is 100 ng/ml in this study.
Conclusion:
GNP-IPCR may provide a highly sensitive method for the detection of FMDV.
Insights
A new gold nanoparticle improved immuno-PCR (GNP-IPCR) assay offers highly sensitive detection of Foot-and-mouth disease virus (FMDV). This method significantly outperforms traditional ELISA for identifying FMDV in clinical samples.
Area of Science:
- Veterinary Virology
- Immunodiagnostics
- Nanotechnology in Diagnostics
Background:
- Foot-and-mouth disease (FMD) is a highly contagious viral disease affecting artiodactyls, posing significant epidemic risks.
- Long-distance transmission and rapid spread necessitate sensitive and specific FMD virus (FMDV) detection methods.
- Existing diagnostic techniques require improvement for early and accurate FMDV identification in clinical settings.
Purpose of the Study:
- To evaluate the efficacy of a gold nanoparticle (GNP) improved immuno-PCR (GNP-IPCR) assay for FMDV detection.
- To assess the sensitivity and specificity of GNP-IPCR compared to conventional methods.
- To determine the utility of GNP-IPCR for analyzing clinical FMDV samples.
Main Methods:
- Development of a GNP-IPCR assay derived from the bio-barcode assay (BCA) technique.
- Utilized polyclonal antibodies for FMDV capture on ELISA microplates.
- Employed GNPs modified with oligonucleotides and a specific monoclonal antibody (MAb 1D11) for signal amplification, followed by PCR and real-time PCR.
Main Results:
- The GNP-IPCR assay demonstrated a detection limit of 10 fg/ml for purified FMDV particles.
- Achieved high sensitivity in detecting FMDV in clinical samples.
- Conventional ELISA showed a detection limit of 100 ng/ml in the same study.
Conclusions:
- GNP-IPCR offers a highly sensitive diagnostic method for FMDV detection.
- The assay's sensitivity surpasses conventional ELISA, enabling earlier and more accurate diagnosis.
- GNP-IPCR holds promise for improved FMDV surveillance and control strategies.
