酶联免疫吸附试验检测新蝶呤的方法学评价

Evaluation of ELISA Method for Detection of Neopterin

目的对应用酶联免疫吸附试验（ELISA）检测新蝶呤（neopterin，Npt）进行方法学评价，同时探讨其在病毒性脑炎患儿脑脊液中的表达水平。方法对ELISA法测定Npt进行灵敏度、精密度、回收率、干扰试验、正常参考值进行检测，并采用该ELISA法检测50例健康人外周血、30例病毒性脑炎患儿脑脊液和12例对照组脑脊液中Npt含量。结果该方法的检测灵敏度为1．08nmol／L，批内精密度为4．94％～5．55％，批间精密度为5．99％～6．57％，回收率为95．8％～107．8％，抗干扰性符合临床要求，健康人血清正常参考值为0～7．84nmol／L，病毒性脑炎患儿脑脊液中Npt含量为（34．09±36．34）nmol／L，对照组脑脊液中Npt含量为（4．55±2．89）nmol／L，病毒性脑炎患者脑脊液中Npt含量明显高于对照组（P〈0．01）。结论ELISA法可用于临床检测血清或脑脊液Npt，适合各级医院临床开展使用。在病毒性脑炎患者的诊断和疗效观察方面有一定价值。

OBJECTIVE To evaluate the detect method of neopterin（Npt） by enzyme-linked immunosorbent assay （ELISA） and discuss the application of Npt in the viral encephalitis. METHODS The Npt was detected by ELISA. The methodology was investigated by sensitivity, precision, rate of recovery, interference and reference value. The apt was detected by ELISA assay in peripheral blood of 50 healthy people, 30 cases of cerebrospinal fluid in children with viral encephalitis and 12 cases of cerebrospinal fluid in the control group. RESULTS In this method, within-run CVs were 4.94% and 5.55% between-run CVs were 5.99% and 6.57%. The sensitivity was 1.08 nmol/L; the rate of recovery was 95. 8- 107. 8M. Various indexes of the methodology coincided with the requirements of clinical laboratory. The reference value of serum Npt was 0- 7. 84 nmol/L. The Npt of cerebrospinal fluid was （34.09 ± 36.34） nmol/L in the viral encephalitis group, and （4.55 ± 2.89）nmol/L in the control group, and the Npt of cerebrospinal fluid in patient was significantly higher than that in control group （P 〈0.01）. CONCLUSIONS ELISA is a valuable method for determination of Npt in the cerebrospinal fluid or serum, which could be applicable in hospitals. Npt may be an index of diagnosis and cure effect detection of viral encephalitis.