2006 Fiscal Year Final Research Report Summary
Development of highly sensitive and versatile methods for evaluation of food safety
| Project/Area Number |
16380091
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Food science
|
|---|
| Research Institution | KYUSHU UNIVERCITY |
|---|
Principal Investigator |
MATSUMOTO Kiyoshi Kyushu University, Faculty of Agriculture, Professor, 大学院農学研究院, 教授 (80038322)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MIYAMOTO Takahisa Kyushu University, Faculty of Agriculture, Professor, 大学院農学研究院, 教授 (70190816)
MATSUI Toshiro Kyushu University, Faculty of Agriculture, Associate Professor, 大学院農学研究院, 助教授 (20238942)
|
|---|
| Project Period (FY) |
2004 – 2006
|
| Keywords | Immuno-PCR / Bisphenol A / Nonylphenol / Surface plasmon resonance / Allergen / Bacillus / RAPD / Emetic Bacillus cereus |
|---|
| Research Abstract |
1. A method for the determination of bisphenol A (BPA), a representative endocrine-disrupting chemical, was developed using a surface plasmon resonance (SPR) sensor. BPA determinations were performed by an indirect competitive immunoassay. The lowest detection limit for BPA by SPR was 1 ppb.
2. An antibody for 4-nonylphenol (NP), which is suspected to be an endocrine disrupting compound, was prepared and used to detect NP. The antibody showed stronger affinity with NP having branched alkyl-group than with NP having straight chain. NP was detected at 0.5 ppm level by an indirect competitive immunoassay with a SPR method.
3. Real time immuno-PCR methods were performed for the detection of ovalbumin (OVA) and lysozyme. An anti-mouse IgG antibody immobilized on magnetic beads were used for the immuno-reaction. A reporter DNA from pBluescript was amplified and used. OVA and lysozyme were detected at 0.1 ppb and 0.1 ppm level, respectively.
4. The Bacillus species and related genera isolated fr
… More
om food samples and identified by using the commercially available biochemical screening kit were analyzed by RAPD analysis using the selected primer combined with morphological observation of colonies, vegetative cells and spores. By the RAPD analyses of the template DNA from bacterial colonies, B. brevis, B. circulans, B. firmus, B. licheniformis, Paenibacilluus macerans, P. polumyxa, B. pumilus, B. sphaericus, B. subtilus and B. thuringiensis were identified by the combination of the present RAPD analysis and morphological observations.
5. A PCR-based method to differentiate between Bacillus cereus and B. thuringiensis and to detect B. cereus strains which cause the emetic type of food-poisoning was developed. 49 strains of B. cereus, isolated from dairy products and processing factories were characterized, and the PCR assay was applied for the detection of these strains.
6. A rapid and simultaneous detection method for 6 main food poisoning bacteria was developed by using genomic DNA of 6 species cultured in SEB medium for 18 hrs at 37C. 6 main food-poisoning bacteria were simultaneously detected in only two reaction-tubes by a multiplex real-time PCR with 5'-nuclease method using target-specific primer sets and fluorescent labeled probes. Less
|
