Strategies for upgrading analyte detection in immuno-PCR studied on identification of type A botulinum neurotoxin
| Autor: | Alexander V. Kolesnikov, A. V. Kozyr, I. G. Shemyakin, A. K. Ryabko, I. V. Zharnikova, A. E. Khlyntseva |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Analyte Chromatography biology Chemistry NeutrAvidin Applied Microbiology and Biotechnology Biochemistry Molecular biology law.invention Matrix (chemical analysis) 03 medical and health sciences 030104 developmental biology Real-time polymerase chain reaction law Polyclonal antibodies Biotinylation TaqMan biology.protein Polymerase chain reaction |
| Zdroj: | Applied Biochemistry and Microbiology. 52:110-120 |
| ISSN: | 1608-3024 0003-6838 |
| Popis: | An efficient method of highly sensitive, specific determination of botulinic type A neurotoxin (which allows the detection of a toxin in a concentration from 1 pg/mL (10 fg per one PCR reaction) was created based on the technology of immuno-PCR. Solid phase consisting of paramagnetic organo–silicious particles carrying a detection complex of the toxin heavy chain with a pair of polyclonal antibodies to it was the most appropriate in the optimal scheme of the analysis. One of the antibodies bound to DNA (which is a matrix for PCR amplification) by means of biotin–neutravidin interaction. Matrix DNA was a molecular “net” generated by biotinylated DNA molecules with tetravalent neutravidin. The binding signal was detected by real time PCR (by the TaqMan method). An immuno-PCR method for determination of type A botulotoxin was developed. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink