Detection of Clostridium botulinum neurotoxin type A using immuno-PCR

Citation
Hc. Wu et al., Detection of Clostridium botulinum neurotoxin type A using immuno-PCR, LETT APPL M, 32(5), 2001, pp. 321-325
Citations number
16
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
LETTERS IN APPLIED MICROBIOLOGY
ISSN journal
0266-8254 → ACNP
Volume
32
Issue
5
Year of publication
2001
Pages
321 - 325
Database
ISI
SICI code
0266-8254(200105)32:5<321:DOCBNT>2.0.ZU;2-0
Abstract
Aims: An immuno-polymerase chain reaction (immuno-PCR) has been developed f or the sensitive detection of antigens, which greatly extends the detection limits of immunoassays. In the current study, the method was applied to th e detection of Clostridium botulinum neurotoxin type A (BTx-A). Methods and Results: Anti-BTx-A antibody-DNA conjugates were synthesized us ing a heterobifunctional cross-linker reagent to covalently link the report er DNA and the antibodies. The antibody-DNA conjugates with antigens were a mplified by PCR, and dose-dependent relationships for each analyte were dem onstrated. Detection limits of immuno-PCR for BTx-A (3.33 x 10(-17) mol) ex ceeded the conventional enzyme-linked immunosorbent assay (3.33 x 10(-14) m ol) by a 1000-fold enhancement in detection sensitivity. Conclusions: Detection of BTx-A antigens by immuno-PCR demonstrated 100% se nsitivity and 100% specificity in 100-fold magnitude below the detection li mit of ELISA. Significance and Impact of the Study: It is concluded that the immuno-PCR m ethod could be used to detect a very low level of BTx-A for clinical diagno sis.