The Study On Serum Protein Fingerprinting For The Diagnosis Of Histologic Chorioamnionitis

PartⅠAt present, the protein chip technology is used to detect of protein fractions in serum,and which had got acquired some achievements. Generally considered weak cationicprotein chip is an ideal protein sepaeation techniques. In view of the considerationsof the cost of testing, the bead (also named liquids chip) is expected to replace theprotein chip for protein separation.Objective: By comparing two kinds of the protein fingerprint in serm of a healthyfull-term pregnant women, which were respectively obtained from weak cationexchange beads (MB-WCX) and weak cation chip protein chips (WCX2).Method: Firstly, we separated respectively protein (peptides) fractions of serumwhich was taken from the same healthy full-term pregnant women, and obtainedprotein fingerprint, with weak cation exchange beads (MB-WCX) and weak cationchip protein chips (WCX2). And then compared the two proteins (peptide) that wereceived, by the number of peaks, peak intensity and peak area of mass spectrum.Results: There were147protein (peptide) peaks were found, and separated byMB-WCX magnetic bead, and140protein (peptide) peaks from WCX2, and peakarea and peak intensity of mass spectrum were equivalent. After comparison, the twoseparation effect is similar, but MB-WCX is cheaper than WCX2. Conclusion: In view of the considerations of the cost of testing, MB-WCX wasbetter than WCX2in protein separation, MB-WCX magnetic bead would be used forthe large scale patient’s serum testing.PartⅡObjective: In the study, we in order to detect the serum proteomics in histologicchorioamnionitis and normal controls by using surface enhanced laser desorptionionization-time of flight mass spectrometry (SELDI-TOF-MS) technology couplewith magnetic beads(MB-WCX), and to screen specific protein biomarkers, and toestablish the diagnosis models of the histologic chorioamnionitis, and evaluate itsclinical significance.Method: Firstly, the nanobeads (MB-WCX) was used to detect the serum proteincomponents of106serum samples, which composed of54cases from pregnantwomen with histologic chorioamnionitis and52cases from pregnant women withnormal control; Secondly, the surface enhanced laser desorption ionization-time offlight mass spectrometry (SELDI-TOF-MS) technology was used to screen specificprotein biomarkers, we randomly selected30cases of histologic chorioamnionitisand30cases of normal controls. Thirdly, we searched the differently expressedproteins by biomark wizard software, which was significant difference(P<0.01).Lastly, set up diagnostic patterns of histologic chorioamnionitis by biomarkpatterns software and obtained its positive predictive value and accuracy andspecificity.Results: There were24differential proteins (peptides) in serum between histologicchorioamnionitis and healthy controls, which were significant difference (P<0.01).8peaks (15903Da\32027Da\6434Da\6841Da\16659Da\8564Da\16070Da and 8128Da) were up-regulated,16peaks (1549Da\1574Da\1523Da\1280Da\1056Da\1481.672Da\2957Da\39288Da\15844Da\1255.93Da\4291Da\1033Da\1234Da\4654Da\1168Da and11737Da) were down-regulated. We selected five differentialproteins (6434Da\1481Da\1280Da\15903Da and1523Da) were used to establish adiagnostic model for histologic chorioamnionitis with sensitivity of83.33%andspecificity of96.67%.Conclusion: This study provides a better method for clinical molecular diagnosis ofhistologic chorioamnionitis, and develops a novel clinical assay for histologicchorioamnionitis, which made it possible that early diagnosis of histologicchorioamnionitis, this technique is an efficient and quick approach for discovery ofprotein markers in serum.24protein biomarkers have been found,5have beenidentified, and also were established the diagnostic models for histologicchorioamnionitis. In addition, a method of the purification and identification of smallmolecular weight proteins has been established in this study to provide a platform fordiscovery of biological markers by SELDI-TOF-MS technique, which will gaininsight into the pathogenesis of histologic chorioamnionitis.