Skip to main content

IMIS

A new integrated search interface will become available in the next phase of marineinfo.org.
For the time being, please use IMIS to search available data

 

[ report an error in this record ]basket (1): add | show Print this page

one publication added to basket [391473]
Ultrasensitive quantification of crustacean tropomyosin by immuno-PCR
Radomirovic, M.; Gligorijevic, N.; Stanic-Vucinic, D.; Rajkovic, A.; Cirkovic Velickovic, T.C. (2023). Ultrasensitive quantification of crustacean tropomyosin by immuno-PCR. International Journal of Molecular Sciences 24(20): 15410. https://dx.doi.org/10.3390/ijms242015410
In: International Journal of Molecular Sciences. MDPI AG: Basel. ISSN 1661-6596; e-ISSN 1422-0067, more
Peer reviewed article  

Available in  Authors 

Keywords
    Crustacea [WoRMS]
    Marine/Coastal
Author keywords
    tropomyosin; immuno-PCR; crustacean allergen; ELISA; shellfish allergen; allergen quantification

Authors  Top 
  • Radomirovic, M.
  • Gligorijevic, N.
  • Stanic-Vucinic, D.
  • Rajkovic, A., more
  • Cirkovic Velickovic, T.C., more

Abstract

    Tropomyosin is the major and predominant allergen among shellfish. This study developed an ultrasensitive immuno-PCR method for the quantification of crustacean tropomyosin in foods. The method couples sandwich ELISA with the real-time PCR (rtPCR) amplification of marker DNAs. Monoclonal anti-TPM antibody was the capture antibody, polyclonal rabbit anti-shrimp tropomyosin antibody was the detection antibody, while natural shrimp tropomyosin served as the standard. A double-stranded amino-DNA was covalently conjugated to a secondary anti-rabbit antibody and subsequently amplified and quantified via rtPCR. The quantification sensitivity of immuno-PCR was 20-fold higher than analogous ELISA, with LOQ 19.8 pg/mL. The developed immuno-PCR method is highly specific for the detection of crustacean tropomyosin and is highly precise in a broad concentration range. Tropomyosin recovery in the spiked vegetable soup was 87.7–115.6%. Crustacean tropomyosin was also quantified in commercial food products. The reported immuno-PCR assay is the most sensitive method for the quantification of crustacean tropomyosin and is the first immuno-PCR-based assay for the quantification of food allergen and food protein in general. The described method could be easily adapted for the specific and ultrasensitive immuno-PCR-based detection of traces of any food allergen that is currently being quantified with ELISA, which is of critical importance for people with food allergies.


All data in the Integrated Marine Information System (IMIS) is subject to the VLIZ privacy policy Top | Authors