Design of a chimaeric antigen and its use in the detection of IgG antibodies against rubella virus

Virology Journal

Xin Xue¹  Ruiping Hu¹  Li Li¹  Hong Yuan¹  Chunli Ma¹  Shumei Ye¹  Jingnan Zhang¹  Huiting Xue¹  Yanhua Ma²  Wenyue Xing³ 

[1] College of Basic Medicine, Inner Mongolia Medical University, 010110, Hohhot, Inner Mongolia, China;College of Basic Medicine, Inner Mongolia Medical University, 010110, Hohhot, Inner Mongolia, China;School of Life Sciences, Inner Mongolia University, Hohhot, China;College of Basic Medicine, Inner Mongolia Medical University, 010110, Hohhot, Inner Mongolia, China;The People’s Hospital of Jiang County, 043600, Shanxi, China;

关键词: Rubella virus;    Prokaryotic expression;    Serological diagnosis;    Chromatography purification;   

DOI  :  10.1186/s12985-022-01760-y

来源: Springer

【 摘 要 】

BackgroundRubella virus (RV) is the causative agent of rubella or German measles. Although most infections cause only mild self-limited measles-like illness, the infection in pregnant women can cause severe foetal malformation or even miscarriage, especially in the first 3 months of pregnancy. Therefore, it is of great practical significance to establish a simple and sensitive RV detection method.MethodsThe partial epitopes of the E1 and E2 proteins from Rubella Virus were selected as the target sites, the sequence of the selected antigenic sites of the E1 and E2 were linked by a linker. The expression plasmid P6T was constructed by inserting the gene into PET-32A + with a histidine Tag. The P6 protein was induced and expressed in Escherichia coli L21 (DE3) and purified by nickel column affinity. The protein P6 antigen was identified by Western blotting analysis, and an anti-P6 antibody ELISA was established to test known serum samples to evaluate the capability of this method.ResultsAfter purification, the concentration and purity of the protein P6 were 0.283 mg/mL and more than 80%, respectively. Western blotting analysis showed that the protein P6 could react with rubella virus positive serum. By ELISA, 36 negative sera and 58 positive sera were detected. The coincidence rate, specificity and sensitivity of the ELISA were 86.2%, 88.89% and 84.48%, respectively. The P6 ELISA with a kappa coefficient of 0.715, P < 0.05, indicated excellent consistency.ConclusionsThe protein P6 with excellent antigenicity obtained from prokaryotic expression followed by chromatography purification could prove useful for early diagnosis of RV infection.

【 授权许可】

CC BY