Detection Of Platelete-associated Antibodies On Megakaryocytes’ Surface For Diagnosis Of Thrombocytopenia

【Objective】 To detect the platelete-associatedimmunoglublin (PAIg) of patients with immune and nonimmunethrombocytopenia by immunofluorescence(FIT), and toevalue the clinica value of detecting platelete-associatedimmunoglublin (PAIg) with immunofluorescence(FIT) by which wecan diagnoge and antidiastole the immune and non immunethrombocytopenia.【Objects and Methods】 we extract serum from30patients whowere clinically diagnosed with immune thrombocytopenia,30cases of patients were non immune thrombocytopenia, and15cases of healthy adult volunteers. All objects were fromhematological medical department in affiliated hospital ofLuzhou Medical College. They were divided into immunity group,non immune group and the normal group. We subculturedmegakaryocyte cell lines (purchased from Shanghai center ofdetection of the assay and ELISA kit) into megakaryocytes,then incubated the serum of three groups and the normalmegakaryocytes which were cultured befroe, Megakaryocytes which were cultured before were smeared, fixed, covered with1%calf serum albumin (BSA), incubated, and then closed atroom temperature, after that, adding and incubating the testedserum on the slide, incubating both FITc labeled mouseanti-human CD61(MAH-CD61, FITc) and Dezhou red labeled Goatanti-human immunoglobulin G(GAH-IgG-Texas Red), we washed anddryed them the next morning. We observe the megakaryocytes’surface fluorescence color under the fluorescence microscope,and analyse GAH-IgG-Texas Red fluorescence intensity level,this make us compared the difference in serum PAIgG levelsbetween the three groups and analyse them.【Results】1. According to the formula,IFT was on qualitativedetection of immune thrombocytopenic’s PAIgG whose deagnosticsensitivity was90.0%, specificity was66.7%, positivepredictive value was72.9%, negative predictive value was86.9%; the rate of misdiagnosis was33.3%, the rate of misseddiagnosis was10.0%, diagnostic efficiency was78.3%. Thesensitivity of quantitative detection was93.3%; specificitywas63.3%, positive predictive value was71.8%, negativepredictive value was90.5%; the rate of misdiagnosis was36.7%,the rate of missed diagnosis was6.7%, diagnostic efficiencywas78.3%.2.PAIgG levels in the immune and non-immune groups were higher than normal group, specially, the group of immunethrombocytopenia increased more obviously.【Conclusion】:1、We detect PAIgG by IFT whose sensitivity andnegative predictive value was high, it could be used asscreening indicators for those patients when they was firstdiagnosed as thrombocytopenia, thrombocytopenia can beexcluded preliminarily if the result was negative, but thepositive results were still not used as a basis for diagnosis,the diagnosis should be excluded by further examinations thatreduced the rate of clinical misdiagnosis and missde diagnosis.2、In view of the platelet’s activation and aggregation beingeasy and the difficult problem of getting normal peoples’platelet, in this experiment we cultured megakaryocyte, anddetected PAIgG of the normal human megakaryocytes’ andthrombocytopenia patients’ serum after incubation by IFT,according to the fluorescence gray value on the megakaryocytecells’ surface, we analysed whether antibody content inimmune and non-immune thrombocy topenia was statisticalsignificance, the more intuitive results excluded theinterference from other antibodies in blood components, so theresult was more reliable.3、For thrombocytopenia, IFT wasexpected to replace the expensive flow cytometry and experimental examination of MAIPA(monoclonal antibodiesspecific captured platelet antigen test) because of its lowprice, which can be spread in our country.4、The diagnosticefficiency was no statistical significance between aualitativeimmunofluorescence and quantitative immunofluorescence,because the former was more rapid, relatively low cost, andmuch more feasibility in clinical, so IFT was more easy to bepromoted.5、 the levels of serum PAIgG in immunethrombocytopenia and non-immune thrombocytopenia was higherthan normal peoples’, immune thrombocytopenia was moresignificant, the degree of difference between immune andnon-immune thrombocytopenia was statistical significance.6、whether to increase the specificity of immune thrombocytopeniabe further studide by combined detection of platelet membraneglycoprotein-specific antibodies.7、Since the detection ofPAIgG have higher sensitivity in this experiment, it can notbe used as identification between primary and secondary immunethrombocytopenia.