title: Evaluation of the clinical value of ELISA based on MPT64 antibody aptamer for serological diagnosis of pulmonary tuberculosis.
abstract: BACKGROUND
  Presently, tuberculosis (TB) poses a global threat to human health. The development of reliable laboratory tools is vital to the diagnosis and treatment of TB. MPT64, a protein secreted by Mycobacterium tuberculosis complex, is highly specific for TB, making antibody to MPT64 a reagent specific for the diagnosis of TB.
  
  
  METHODS
  Antibody to MPT64 was obtained by a combination of genetic engineering and immunization by the system evolution of ligands by exponential enrichment. A high-affinity aptamer of antibody to MPT64 was selected from a random single-stranded DNA library, and a sandwich ELISA method based on this aptamer was developed. This ELISA method was used to detect TB in 328 serum samples, 160 from patients with pulmonary TB (PTB) and 168 from non-tuberculous controls.
  
  
  RESULTS
  The minimum limit of detection of the ELISA method was 2.5 mg/L, and its linear range varied from 10 mg/L to 800 mg/L. Its sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and area under the curve, with 95 % confidence intervals, were 64.4 % (56.7 %-71.4 %), 99.4 % (96.7 %-99.9 %), 108.2 (15.3-765.9), 0.350 (0.291-0.442) and 0.819 (0.770-0.868), respectively. No significant difference in sensitivity was observed between sputum smear positive (73/112, 65.2 %) and negative (30/48, 62.5 %) individuals.
  
  
  CONCLUSIONS
  This sandwich ELISA based on an MPT64 antibody aptamer may be useful for the serological diagnosis of PTB, both in sputum smear positive and negative patients.
authors.affiliation: Department of Medical Laboratory, Changzhou Tumor Hospital Soochow University, Changzhou 213001, China.
authors.forename: Changtai
authors.initials: C
authors.lastname: Zhu
authors.forename: Jinming
authors.initials: J
authors.lastname: Liu
authors.forename: Yang
authors.initials: Y
authors.lastname: Ling
authors.forename: Hua
authors.initials: H
authors.lastname: Yang
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
authors.forename: Ruijuan
authors.initials: R
authors.lastname: Zheng
authors.forename: Lianhua
authors.initials: L
authors.lastname: Qin
authors.forename: Zhongyi
authors.initials: Z
authors.lastname: Hu
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology*; D052157:Aptamers, Nucleotide / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D052156:SELEX Aptamer Technique; D012680:Sensitivity and Specificity; D014397:Tuberculosis, Pulmonary / Q000097:blood / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D052157:Aptamers, Nucleotide; C510537:MPT64 protein, Mycobacterium tuberculosis