酶联免疫吸附试验测定血清苯妥英钠浓度  

Establishment of ELISA for Measuring Phenytoin in Serum

作　　者：苏薇[1] 林其燧[1] 宋耀虹[1] 

机构地区：[1]中国医学科学院

出　　处：《中国医学科学院学报》2000年第1期93-95,共3页Acta Academiae Medicinae Sinicae

摘　　要：目的　建立酶联免疫吸附实验 (EL ISA)测定苯妥英钠血药浓度的方法。方法　采用碳二亚胺法将化学修饰后的苯妥英钠与人血清白蛋白结合 ,再与被氧化的辣根过氧化物酶结合 ,作为酶标抗原 ,自制兔抗苯妥英钠多克隆抗体。结果　抗苯妥英钠 Ig G稀释倍数 1∶ 2 0 0、酶标记抗原 1∶ 10 0时 ,标准曲线最理想 ;该方法的测定灵敏度为 2 .87μg/ ml,线性范围 2 .9～ 30 μg/ m l,批内变异系数 3.3%～ 10 .2 % ,批间变异系数 5 .1%～ 13.2 % ,回收率 90 %～ 96 % ,其它 4种抗癫痫药物 (扑痫酮、苯巴比妥、卡马西平和丙戊酸钠 )对测定无干扰 ,与 HPL C方法相关良好 ,相关系数为 0 .97。结论　该方法具有良好的准确性、精确性和特异性 ,灵敏度和测定范围能够满足临床需要 ,可用于苯妥英钠的临床治疗药物监测。Objective To develop a solid phase,competitive enzyme immunoassay for the measurement of serum phenytoin. Methods The chemical modified phenytoin combined with human serum albumin and then conjugated with horseradish peroxidase (HRP) to produce the enzyme labeled phenytoin(DPH HSA HRP).The anti phenytoin antibody was prepared in this lab. Results The working range and sensitivity of this method were 2 9~30 μg/ml and 2 87 μg/ml respectively.The intro assay coefficient of variation (CV) was 3 3%~10 2% ( n=15 ) and inter assay CV was 5 1%~13 2%.The recovery of this method was 90%~96%.No significant interference was observed with phenobarbital,primidone,carbamazepine,and valproic acid.The new assay method was compared with HPLC method.A linear regression analysis yielded a slope of 1 03,an intercept 1 38 and a correlation coefficient 0 97. Conclusions This solid phase enzyme immunoassay for serum phenytoin appears to be simple,precise,and accurate.It may be readily adopted in clinical laboratory for therapeutic monitoring of phenytoin level in serum. Key words phenytoin;enzyme linked immunoabsorbent assayelative amount of PBP 2a in 10 MRSA were analyzed by SDS PAGE.Statistical signifcance was analyzed with t tests. Results 56 MRSA were isolated from 180 strains staphylococcus aureus through detecting MecA,but only 52 MRSA were idendified using agar plate dilution method and 4 strains were missing for test(MIC of oxacillin 2,2,1,0 5 ug/ml,respectively).In addition,the amount of PBP 2a in 10 MRSA at 37℃,pH5 2 and 9 g/L NaCl were lower than 32℃,pH7 2,and 40 g/L NaCl(P:0 02～0 05,0 01～0 005,0 02～0 05). Conclusion The results showed that temperature,pH and concentration of salt have apparent effects on antimicrobial activities of oxacillin,cefazolin and ceftazidime against MRSA,but the detection of MecA gene may be used to diagnose MRSA infection earlier,without influenced by culture conditions. Key words: protein binding; methicillin resistance; staphy

关 键 词：苯妥英钠 ELISA 测定 抗癫痫药 血药浓度 

分 类 号：R971.6[医药卫生—药品] R969.1[医药卫生—药学]