目的探讨不同储存条件和反复冻融对血清和血浆乙酰胆碱受体抗体(AChRAb)水平的影响。方法将1例重症肌无力患者的血清和血浆标本分成数等份,置于不同储存条件下(常温、4℃或-80℃)并经过不同次数的冻融,不同等分的样本使用同一批次商品化ELISA试剂盒双平行孔检测AChRAb水平。计算各种条件下血清和血浆标本AChRAb抑制率的变异系数(CV),比较不同处理条件下的抑制率以及同一条件下血清和血浆标本的抑制率。结果除常温条件下血浆标本CV〉10%外,其余条件下CV均〈10%。4℃条件下血清AChRAb的抑制率显著高于其他条件下血清的抑制率(P〈0.05),4℃和常温条件下血浆的抑制率显著低于其他条件下血浆标本的抑制率(P〈0.05)。4℃条件下血清和血浆标本AChRAb抑制率差异有统计学意义(P〈0.05),而冷冻保存后经过多达5次冻融两者的抑制率差异无统计学意义(P〉0.05)。结论 -80℃直接储存的血清和血浆标本,即使经过多次冻融,AChRAb水平检测结果仍可较为一致。血清标本在常温、4℃或冷冻保存,血浆标本低温冷冻保存,数日内完成检测结果具有一致性。
Objective To evaluate the intra-assay reproducibility of anti-acetylcholine receptor antibody(AChRAb)level in serum and plasma samples among various pre-treatment conditions consisting of different storage schemes and subsequent freeze/thaw cycles.Methods The serum and plasma samples were collected from a patient with myasthenia gravis and allocated in aliquots pre-treated in various conditions(stored at room temperature[RT],4℃ or-80℃ with subsequent freeze/thaw cycles).Each aliquot was examined in duplicate with commercial AChRAb ELISA kit.The coefficient of variability(CV)was calculated for inhibition rate of AChRAb in each pre-treatment condition.Inhibition rates of AChRAb in serum or plasma samples were compared among each condition,and compared between serum and plasma samples in each condition.Results Except for plasma sample at RT,the CVs of all the other samples in various conditions were10%.The inhibition rate of AChRAb in serum sample was significantly higher at 4℃ than those in other conditions(P 0.05).The inhibition rate in plasma samples were significantly higher at 4℃ and RT than those in other conditions(P〈0.05).There were significant difference in inhibition rates between serum and plasma samples at 4℃(P〈0.05),while no difference was found in frozen conditions with up to five subsequent freeze/thaw cycles(P〈0.05).Conclusions There was good intra-test reproducibility of the ELISA kit in determining the AChRAb levels in serum and plasma samples stored at-80℃ directly with various subsequent freeze/thaw cycles.There was good intra-test reproducibility in serum samples stored at RT,4℃ and frozen conditions and in plasma samples stored in frozen conditions,which were examined in several days after blood collection.