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A Novel Menthod for the Rapid Detection of Porcine Streptecoccal Disease Based on Quantum Dot Fluorescent Probes

Author: WuHongMin
Tutor: HanHeYou
School: Huazhong Agricultural University
Course: Pesticides
Keywords: Quantum dot fluorescent probes Streptococcus suis type 2 Muramidase released protein Rabbit anti -pig IgG Fluorescence Immunoassay High-throughput detection
CLC: S858.28
Type: Master's thesis
Year: 2009
Downloads: 61
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Abstract


Streptococcus suis is a zoonotic disease caused by Streptococcus suis (SS), 35 serotypes of Streptococcus suis type 2 (SS2) pop the most extensive, the most pathogenic, so the development and establishment of SS2 important virulence factor - muramidase released protein (MRP) swine streptococcus disease rapid detection method for the detection and diagnosis of the disease is of great significance. Quantum dots as a novel fluorescent probe, has many unique advantages in the optical, electrical, chemical, chemistry, materials science, biomedical and environmental detection aspects has become an important research tool. This paper combines fluorescence immunoassay technology using quantum dot-MRP antibody protein probes and quantum dots - rabbit anti-pig IgG probe detection of Streptococcus suis, main contents and results are as follows: 1. Synthesis of thioglycolic acid modified CdSe / ZnS core-shell quantum dots and CdSe / ZnS quantum dot-MRP antibody protein probes were prepared using polyacrylamide gel electrophoresis and molecular spectroscopy to study the bonding mechanism of MRP antibody protein with CdSe / ZnS quantum dots, the results that MRP antibody protein with CdSe / ZnS quantum dots may be combined by surface coordination role. The fluorescence spectrometry Optimization of CdSe / ZnS quantum dot-MRP antibody protein probes prepared influencing factors, to establish a new method for the determination of the MRP antigen protein, its linear range of 5.0 × 10 -8 ~ 1.5 × 10 -6 mol / L, the linear correlation coefficient of 0.998. 2. synthesis of glutathione-modified water-soluble CdTe quantum dots, and EDC · HCl / Sulfo-NHS The covalent coupling agents prepared MRP antibody protein-based quantum dot fluorescent probes. Molecular spectroscopy, capillary electrophoresis, transmission electron microscopy and fluorescence microscopy probe characterization, the results show that the quantum dot probes maintain MRP antibody protein's biological activity and biological specificity. Fluorescence immunoassay MPR antigen protein detection, having a good linear relationship with the fluorescence intensity and the concentration of antigen, and a linear concentration range of 5.0 × 10 -8 1.5 × 10 - 5 mol / L. Compared with the plate count method, the test of quantum dot probes for the detection of pig offal serum obtained satisfactory results. The same time, the test method in 96-well plates for testing and to provide important reference for the realization of high-throughput detection of Streptococcus suis. Prepared CdTe quantum dot probes rabbit anti-pig IgG-based, and for the detection of MRP antigen protein and serum of pig offal, the results show that this method of concentration 2.5 × 10 -7 to 1.5 × 10 -5 mol / L range between antigen has a good linear detection relationship. The detection of the sera of pig offal, in this test method and the plate assay and compared based on the quantum point probe method for the detection of the MRP antibody protein to obtain a good consistency. Examined the time stability of the quantum dot probes, the results showed that the preparation of quantum dot probes has good stability, and provide important reference for the preparation of the test kit for Streptococcus suis. In this study, the quantum dot probes used for the research in the field of major animal diseases, not only provides a new, simple and rapid, high-throughput swine streptococcus disease detection, disease detection kit for Streptococcus suis preparation as well as quantum point fluorescent probes provide important reference for other major animal disease research.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Pig
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