An Universal Colloidal Gold Detection Method of Vitellogenin as an Environmental Estrogen Biomarker

Vitellogenin is a kind of indicator of environmental estrogen or environmental estrogenic mimics. The development of a colloidal gold immunochromatography method for rapid detection of vitellogenin can make a certain contribution to the detection of estrogen pollution in the ecological environment. In this experiment, we established a colloidal gold immunochromatography test strip detection method by preparing monoclonal antibodies against vitellogenin. First of all, we used the vitellogenin of the Bostrychus sinensis as the antigen obtained in the laboratory previously and injected it into laboratory mice to obtain the monoclonal antibody against the vitellogenin. Then, the colloidal gold particles were prepared by sodium citrate reduction method, and prepared monoclonal antibody was used as the labeled antibody to assemble the colloidal gold immunochromatographic test strip. This method was used to test the standards of different dilutions, and it was found that when the dilution reached 1:2000, obvious bands could also be observed on the T line. In addition, this method was also used to detect the serum of aquatic species such as Siniperca chuatsi , Oryzias latipes , Bostrychus sinensis , Abalone , Procambarus clarkii , ♀ Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatus and Hapalogenys kishinouyei . After the experiments of RNA extraction, cDNA reverse transcription, and PCR on the hepatopancrease of the Procambarus clarkii and the liver of the ♀ Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatus , Gobiocypris rarus and Hapalogenys kishinouyei , the PCR products were sent to Sangon Biotech Company for further sequencing. The sequencing results were compared with the vitellogenin sequence of Bostrychus sinensis . The comparison revealed that there is a certain length of the same protein sequence among the above five kinds of species.