title: Development of a novel anti ESAT-6 monoclonal antibody for screening of Mycobacterium tuberculosis.
abstract: Tuberculosis (TB) is a major global public health problem with an estimated one-third of the world's population infected with Mycobacterium tuberculosis, thus necessitating fast and accurate diagnosis of TB. However, the diagnosis of latent infection remains difficult due to the lack of a simple, reliable test for M. tuberculosis infection. The objective of the current study was to generate and characterize a novel monoclonal antibody (mAb) that specifically target the early secretory antigenic target 6 (ESAT-6) protein for tuberculosis (TB) immunological diagnosis. The BALB/c mice were immunized by a peptide with 13 amino acids (amino acid residues 24 to 36) of ESAT-6 protein. Stable hybridomas cell lines were established and mAb was identified by Western Blot, immunoprecipitation (IP) and Enzyme linked immunosorbent assay (ELISA). In addition, this mouse mAb was used to coat plates, and biotin-labelled polyclonal antibodies were used to detect the antigens. Finally, the antibody was verified in 280 patient sputum culture supernatants and pleural effusion aspirates. The positive detection rate of the generated ESAT-6 mAb was 91% in the clinical diagnosis of tuberculosis pleural effusion, and this mAb had a sensitivity of 92.4% and a specificity of 100% in all M. tuberculosis cases. Our data indicated that the mAb generated in this study can potentially serve as a tool in the laboratory diagnosis of TB.
authors.affiliation: Central Laboratory, Hangzhou First People's Hospital (Affiliated Hangzhou Hospital, Nanjing Medical University) Hangzhou 310006, P.R. China.
authors.forename: Jianhang
authors.initials: J
authors.lastname: Leng
authors.affiliation: Central Laboratory, Hangzhou First People's Hospital (Affiliated Hangzhou Hospital, Nanjing Medical University) Hangzhou 310006, P.R. China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Deparment of Endocrinology, Red Cross Hospital of Hangzhou Hangzhou 310003, P.R. China.
authors.forename: Chengmin
authors.initials: C
authors.lastname: Shou
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine Hangzhou 310003, P.R. China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Central Laboratory, Hangzhou First People's Hospital (Affiliated Hangzhou Hospital, Nanjing Medical University) Hangzhou 310006, P.R. China.
authors.forename: Guangchao
authors.initials: G
authors.lastname: Zhuo
authors.affiliation: Central Laboratory, Hangzhou First People's Hospital (Affiliated Hangzhou Hospital, Nanjing Medical University) Hangzhou 310006, P.R. China.
authors.forename: Keyi
authors.initials: K
authors.lastname: Wang
authors.affiliation: Central Laboratory, Hangzhou First People's Hospital (Affiliated Hangzhou Hospital, Nanjing Medical University) Hangzhou 310006, P.R. China.
authors.forename: Junya
authors.initials: J
authors.lastname: Shen
authors.affiliation: Deparment of Pneumology, Hangzhou First People's Hospital (Affiliated Hangzhou Hospital, Nanjing Medical University) Hangzhou 310006, P.R. China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Huang
keywords: Anti-ESAT-6; Mycobacterium tuberculosis; immunological detection; monoclonal antibody