| Purposes:(1)screen a suitable protein chip from two kinds of chips CM10 and IMAC-3-Cu.(2)To discover the protein(s)with relation to tuberculosis and gastric carcinoma and provide new ideas of finding new biomakers of tuberculosis and gastric carcinoma,while establish diagnostic models of tuberculosis and gastric carcinoma and investigate its feasibility of the clinical application;Methods:(1)Serum samples from 8 cases of tuberculosis,8 cases of gastric carcinoma and 8 cases of normal control people were collected respectively, special protein or peptide spectra were detected by SELDI measurement after treating the samples onto two kinds of protein chips CM10 and IMAC-3-Cu for each case.According to the protein profiling results,screening suitable protein chips.(2)Serum samples were collected from 58 cases of tuberculosis(including 6 cases of multiple-drug-resistant tuberculosis),42 cases of gastric carcinoma and 53 cases of normal people.The samples were profiled by SELDI measurement with CM10 chips offered by Ciphergen Corporation.Ciphergen Biosystems softwares were used to analyze the proteomic profiles.Results:(1)Analysis of serum protein fingerprints showed CM10 chip combined more proteins and more which show higher intensity in peaks than IMAC-3-Cu chip.(2)Comparison of the group of tuberculosis and the group of normal people:analyze two groups of 60 samples and get 133 protein peaks,among them 86 show the difference obviously in the two groups.Compared with the normal people group,42 proteins were up-regulated expressed and 44 down-regulated expressed.Make use of those differentiated proteins,a model was built.Accurate rate of this model was 98.33%,sensitivity 96.67%, specificity 100%,positive predictive value 100%(29/29),negative predictive value 96.77(30/31),and Az in ROC in it 0.983.The 105 unknown serum samples were measured and predicted while mass spectra of the serum proteins of each sample were navigated into the model for blind test,accurate rate of which is 83.81%(88/105),sensitivity 71.15%(37/52),specificity96.23% (51/53),positive predictive value 94.87%(37/39),negative predictive value 77.27%(51/66),and Az in ROC in it 0.837.(3)Comparison of the group of multiple-drug-resistant tuberculosis and the group of normal people:two groups of 36 samples were detected and 160 proteins peaks were attained,among them 37 show the difference obviously in the two groups.Compared with the group of normal people,9 proteins were up-regulated expressed and 28 proteins down-regulated expressed.Make use of those differentiated proteins,a model was achieved,accurate rate of which was 100%(36/36),sensitivity 100%(6/6),specificity 100%(30/30),positive predictive value 100%(6/6),negative predictive value 100%(30/30),and Az in ROC in it 1.000.(4)Comparison of the group of multiple-drug-resistant tuberculosis and the group of tuberculosis:two groups of 36 samples were detected and 110 proteins peaks were acquired,among them 25 show the difference obviously in the two groups.Compared with the group of tuberculosis,6 proteins were up-regulated expressed and 19 proteins down-regulated expressed.Make use of those differentiated proteins,a model was obtained,accurate rate of which was 94.44% (34/36),sensitivity 83.33%(5/6),specificity 96.67%(29/30),positive predictive value 83.33%(5/6),negative predictive value 96.67%(29/30),and Az in ROC in it 0.900.(5)Comparison of the group of gastric carcinoma and the group of normal people:two groups of 60 samples were detected and 151 proteins peaks were obtained,among them 53 show the difference obviously in the two groups. Compared with the normal people group,36 proteins were up-regulated expressed and 17 proteins down-regulated expressed.Make use of those differenced proteins,a model was attained,accurate rate of which was 90% (54/60),sensitivity 80%(24/30),specificity 100%(30/30),positive predictive value 100%(24/24),negative predictive value 83.33%(30/36),and Az in ROC in it 0.900.The 64 unknown serum samples were measured and predicted while mass spectra of the serum proteins of each sample were navigated into the model for blind test,accurate rate of which was 96.23%(77/95),sensitivity 61.91%(26/42),specificity 96.23%(51/53),positive predictive value 92.86% (26/28),negative predictive value 76.12%(51/67),and Az in ROC in it 0.791.Conclusions:(1)CM10 chip can ottain more significant differentiated protein peaks which can distinguish the tuberculosis patients and gastric carcinoma from the health people compared to IMAC-3-Cu chip.more significant differentiated serum protein of tuberculosis and gastric carcinoma would likely be attained when research of the protein fingerprints was actualized with CM10 chip.(2)There is significant difference among the group of tuberculosis,gastric carcinoma and normal people in serum proteome,and those different proteins are potentially the proteins related to tuberculosis and gastric carcinoma.(3)The technology of proteinchip-SELDI-TOF-MS shows great excellence which is minor sample-dosage depended,operated simply,high in throughput, sencitivity and speciality and great quality in repeated test.And it displays promising and potential detection of tuberculosis and gastric carcinoma in the clinical medicine. |
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