Differential comparison of serum proteome between tuberculosis patients and healthy controls by using two-dimensional polyacrylamide gel electophoresis

Objective To investigate the difference of serum proteome between tuberculosis patients and healthy controls and to screen out novel laboratory diagnosis marker of tuberculosis.Methods Samples of peripheral blood were collected from patients with bacterial positive pulmonary tuberculosis(+group),patients with bacterial negative pulmonary tuberculosis(-group),patients with ex-pulmonary tuberculosis(ex-pulmonary group),healthy controls(control group),with 10 patients in each group.Serum samples of each group were mixed respectively by different proportion according to the protein concentration of the samples.Two-dimensional polyacrylamide gel electrophoresis(2-DE) was performed on immobilized pH gradient rod gels(pH4～7,17 cm) and each mixed sample was tested three times simultaneously.The gels were stained using colloidal Coomassie Blue,and scanned using Umax powerlook2000 scanner to acquire images of 2-DE gel in the condition of projection mode and 300dpi resolution.Analysis of the image was performed with ImageMaster 2D Platium5.0.Results 310±64,307±33,296±54 and 267±23 of clear protein spots were detected respectively in the 2-DE gel of control group,+group,-group and ex-pulmonary group.The matching rates with control group matched+group and-group and ex-pulmonary group,were 74%±2%,76%±2% and 68%±5% respectively.Making a comparison among four groups,there were 21,18,and 24 differentially expressed proteins respectively between control group and+group,-group,ex-pulmonary group.And there were 9,8,7 up-regulation expression proteins in+group,-group and ex-pulmonary group.There were 3 differentially expressed proteins including 2 up-regulation expression proteins in tuberculosis group based on+group and-group and ex-pulmonary group.Conclusion There are differentially expressed proteins in serum between tuberculosis patients and healthy controls and that novel laboratory diagnosis markers of tuberculosis from serum may be found.In this study,primary model of differentially expressed proteins in serum between tuberculosis patients and healthy controls is set,but needs further test.These differential proteins should be identified.