流式微球芯片俘获技术对免疫性血小板减少症患者抗体的检测及临床意义

Detection of antibody in patients with immune thrombocytopenia by flow cytometric immuno-bead array and the clinical significance

目的使用流式微球检测法测定免疫性血小板减少症(ITP)患者自身抗体的表达,探索该方法在诊断ITP患者中的临床意义,并探究血小板特异性自身抗体的种类与临床用药疗效之间的关系。方法对81例ITP患者的抗血小板膜糖蛋白Ⅰb(GPⅠb)抗体、抗血小板膜糖蛋白Ⅱb/Ⅲa(GPⅡb/Ⅲa)抗体表达使用流式微球芯片俘获技术(FICBA)进行检测,将其中71例结果与使用单克隆抗体俘获特异性血小板抗原(MAIPA)法得出的实验结果进行比较。ITP患者分为激素治疗组和重组人血小板生成素(rhTPO)治疗组,两组患者分别为49、28例,其他为4例。另外,依据临床疗效分为有效组和无效组。结果 (1) FICBA法检测结果:抗GPⅠb抗体阳性患者和抗GPⅡb/Ⅲa抗体阳性患者分别为30、29例,阳性率分别为42.25%、40.85%,两种抗体联合检测阳性患者42例,总阳性率59.15%。(2) MAIPA法检测结果:抗GPⅠb抗体阳性患者和抗GPⅡb/Ⅲa抗体阳性患者分别为19、22例,阳性率分别为26.76%、30.99%,两种抗体联合检测阳性患者33例,总阳性率46.48%。(3)比较FICBA法和MAPIA法的实验结果,差异无统计学意义(P>0.05)。(4)对采用激素治疗的49例患者,使用FICBA法检测其抗体表达水平,结果显示抗GPⅠb抗体阳性患者和抗GPⅠb抗体阴性患者分别为19、30例,治疗后的有效率分别为42.11%、73.33%,差异有统计学意义(P<0.05)。结论在血小板特异性自身抗体检测方面,相较于MAIPA法,FICBA法更方便快捷,其敏感度差异无统计学意义,这对ITP患者的临床诊断具有重要的意义。使用糖皮质激素治疗效果不明显的影响因素之一可能是抗GPⅠb抗体阳性。

Objective To detect the expression of autoantibodies in patients with immune thrombocytopenia(ITP) by flow cytometric immuno-bead array, as well as explore its clinical significance in the diagnosis of ITP patients, and the relationship between the types of platelet specific autoantibodies and the clinical effects of drugs. Methods Flow cytometry immuno-bead array was used to detect the expression of anti platelet membrane glycoprotein Ⅰb(GPⅠb) antibody and antiplatelet membrane glycoprotein Ⅱb/Ⅲa(GPⅡb/Ⅲa) antibody. The results of 71 cases were compared with those of monoclonal antibody capture specific platelet antigen(MAIPA). Patients with ITP were halved into the groups of hormone treatment and recombinant human thrombopoietin( rh TPO) treatment,with49 and 28 cases respectively,and 4 cases from the other groups. Moreover,the patients were divided into effective group and ineffective group based on the clinical effects. Results(1) Results of flow cytometric immuno-bead array: There were comparatively 30 and 29 patients with positive anti GPⅠb antibody and anti GPⅡb/Ⅲa antibody,making the positive rates 42. 25% and 40. 85%. Upon the combined detection of the two antibodies,42 patients were finally positive,with a total positive rate of 59. 15%.(2) Results of MAIPA method: there were 19 and 22 cases of patients with the two antibodies,and the positive rates were 26. 76% and 30. 99% respectively. The combined detection of the two antibodies was 33 cases creating a total positive rate of 46. 48%.(3) The comparison between the above two methods showed no significant difference( P > 0. 05).(4) The results of the 49 patients in hormone therapy whose antibody expression was detected by flow cytometry showed that there were 19 and 30 cases with positive anti GPⅠb and with negative anti GPⅠb with the effective rates 42. 11% and 73. 33%. The difference was statistically significant( P < 0. 05). Conclusion Compared with MAIPA method,flow cytometric immuno-bead array is more speedy in detecting the platelet-specific autoantibody,but the sensitivity is not different. It is of great significance for the clinical diagnosis of ITP patients. In addition,the reason why the effect of glucocorticoid treatment is not obvious may be related to the positive anti GPIB antibody.