Development of an Immuno-PCR Protocol for Detection of a Small Amount of Antigen / 대한진단검사의학회지
The Korean Journal of Laboratory Medicine
;
: 66-70, 2005.
Article
Dans Coréen
| WPRIM
| ID: wpr-190285
ABSTRACT
BACKGROUND:
Immuno-PCR has been known as a highly sensitive and specific method, yet no standardized protocol is available. We analyzed each step of immuno-PCR to develop a reliable standardized method.METHODS:
We made a protocol modified from several methods reported previously, and performed immuno-PCR, but false positive reactions were noted. To reduce the false positivity, we investigated the buffer reagents and biotin-labelled oligo-nucleotide probe. Using a finally determined protocol, we compared the detection-limits of the immuno-PCR and ELISA methods.RESULTS:
Streptavidin was identified as a main reagent causing a non-specific binding, thus it was replaced by neutravidin. The employment of CAS block as a dilution buffer for the biotin-labelled oligo-nucleotide probe and Casein block as a buffer for the detection antibodies resulted in a dramatic reduction in the false positive reactions. The standardized immuno-PCR detected angiogenin antigen at a concentration as low as 5 fg/mL, while an ELISA method detected 5 pg/mL.CONCLUSIONS:
The immuno-PCR procedure newly described in this study was ultra-sensitive with no false positivity. This method can be utilized as an epochal tool for detection of a small amount of antigen which would not be discovered by ELISA method.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Test ELISA
/
Caséines
/
Streptavidine
/
Emploi
/
Faux positifs
/
Limite de détection
/
Indicateurs et réactifs
/
Anticorps
Type d'étude:
Etude diagnostique
langue:
Coréen
Texte intégral:
The Korean Journal of Laboratory Medicine
Année:
2005
Type:
Article
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