Cloning expression and serological evaluation on Mycobacterium tuberculosis four new antigens.
10.3760/cma.j.issn.0254-6450.2018.04.026
- Author:
Q LUO
1
;
S J LI
1
;
T Y XIAO
2
;
M C LI
2
;
H C LIU
2
;
Y L LOU
1
;
K L WAN
3
,
4
Author Information
1. School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou 325035, China.
2. State Key Laboratory for Infectious Diseases Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
3. School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou 325035, China
4. State Key Laboratory for Infectious Diseases Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
- Publication Type:Journal Article
- Keywords:
Antigen;
ELISA;
Immunological diagnosis;
Mycobacterium tuberculosis
- MeSH:
Antigens, Bacterial/genetics*;
Cloning, Molecular;
Enzyme-Linked Immunosorbent Assay;
Humans;
Immunoglobulin G;
Mycobacterium tuberculosis/metabolism*;
Polymerase Chain Reaction;
ROC Curve;
Recombinant Proteins;
Sensitivity and Specificity;
Serologic Tests/methods*;
Tuberculosis/genetics*
- From:Chinese Journal of Epidemiology
2018;39(4):514-518
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To evaluate the serological diagnostic value of Mycobacterium (M.) tuberculosis four new antigens Rv0432, Rv0674, Rv1566c and Rv1547. Methods:Rv0432, Rv0674, Rv1566c and Rv1547 were amplified from M. tuberculosis strain H37Rv genomic DNA by using PCR, among which Rv1547 was divided into two segments for amplification (Rv1547-1 and Rv1547-2). The segments were cloned into expression vector PET-32a while the recombinant proteins were purified by affinity chromatography. Serums were incubated with BL21 (DE3) proteins. Antibodies IgG against M. tuberculosis were tested with 151 serum samples (41 healthy people and 110 TB patients) by using ELISA. The diagnostic efficiency of antigens was analyzed by means of receiver operating characteristic curve. Difference of the objective proteins in TB patients and healthy controls was compared by t-test. Results: Recombinant antigens Rv0432, Rv0674, Rv1566c, Rv1547-1 and Rv1547-2 were successfully expressed and purified. Results from ELISA showed that the sensitivity, specificity, positive predictive value, negative predictive value, Youden index and area under the curve of Rv0432, Rv0674, Rv1566c, Rv1547-1 and Rv1547-2, as 43.64%-92.73%, 80.49%-92.68%, 0.92-0.94, 0.38-0.80, 0.363-0.732 and 0.649-0.915. All the objective proteins showed significantly higher antibody levels in TB patients, when compared to the healthy controls (P<0.000 1). Conclusion: The newly identified antigens Rv0432, Rv0674, Rv1566c, Rv1547-1 and Rv1547-2 all performed well when being used for TB serological diagnosis, thus were expected to be new candidate antigens used for TB diagnosis.
