Establishment and application of indirect ELISA for detection of measles virus IgG antibody based on nucleoprotein
Hu Ren, Lei Cao, Naiying Mao, Rongbo Zhang, Peibei Zhao, Hai Li, Ruoqun Ma, Huiling Wang, Yan Zhang, Wenbo Xu
Abstract
ObjectiveTo establish an indirect ELISA method for detection of measles virus antibody based on the nucleoprotein of measles virus expressed in prokaryotic system selected as the coating antigen.
MethodsThe working conditions were screened and optimized to determine the operation process of indirect ELISA. The anti-measles virus IgG antibody in 157 sera of healthy children and mothers of newborn infants was detected and compared with the commercial measles virus ELISA IgG antibody detection kit.
ResultsThe result showed that the coefficient of variation of intra and inter assay repeatability of the indirect ELISA was less than 10%. Compared with the result of the commercial kit, the total concordance rate of serum samples was 95.5%, and the sensitivity and specificity were 94.8% and 98.3%, respectively. There was no significant difference between the two methods(χ2=0.313, P>0.05; χ2=0.000, P>0.05) in the positive rate of measles virus IgG antibody in 85 healthy children at the age of 0~15 years. The level of measles virus antibody detected by ELISA established in this study showed the same increasing and decreasing trend as that detected by the commercial kit in different age groups. The correlation coefficient r of the quantitative result was 0.893 (P< 0.001), which showed the quantitative potential of the method.
ConclusionsThe ELISA method established in this study has good stability, high sensitivity and specificity, and there was no significant difference between the detection results of commercial kit and the ELISA method. It can be used for the seroepidemiological investigation of measles virus and the evaluation of antibody level of the population after vaccination.
Key words:
Measles virus; Nucleoprotein; Serology; Indirect ELISA
Contributor Information
Hu Ren
Medical College, Anhui University of Science and Technology, Huainan 232001, China
Lei Cao
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, WHO WPRO Regional Reference Measles/Rubella Laboratory, Beijing 102206, China
Naiying Mao
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, WHO WPRO Regional Reference Measles/Rubella Laboratory, Beijing 102206, China
Rongbo Zhang
Medical College, Anhui University of Science and Technology, Huainan 232001, China
Peibei Zhao
Handan Center for Disease Control and Prevention, Handan 056004, China
Hai Li
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, WHO WPRO Regional Reference Measles/Rubella Laboratory, Beijing 102206, China
Ruoqun Ma
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, WHO WPRO Regional Reference Measles/Rubella Laboratory, Beijing 102206, China
Huiling Wang
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, WHO WPRO Regional Reference Measles/Rubella Laboratory, Beijing 102206, China
Yan Zhang
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, WHO WPRO Regional Reference Measles/Rubella Laboratory, Beijing 102206, China
Wenbo Xu
Medical College, Anhui University of Science and Technology, Huainan 232001, China
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, WHO WPRO Regional Reference Measles/Rubella Laboratory, Beijing 102206, China
Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China
