CAJ | 학술논문

目的建立检测b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)多聚核糖基核糖醇磷酸盐(polyribosylribitol phosphate,PRP)的竞争酶联免疫吸附法(competative enzyme-linked immunosorbent assay,cELISA).方法以Hib PRP-酪胺(PRP-Ty)为包被抗原,待测PRP为竞争抗原,利用方阵滴定法确定抗原与血清抗PRP抗体的最适反应条件,建立cELISA法并验证其准确性和精密度,并将该法与传统检测法进行比较.结果最适包被PRP-Ty浓度为1.30 mg/L,血清抗PRP抗体稀释度为1∶40 000.该法的检测线性范围为6.25～100.00 mg/L,决定系数(R2)为0.99,批内精密度为3.39％～6.53％,批间精密度为8.48％.该法对Hib PRP的检测结果与传统化学法一致.结论建立的Hib多糖cELISA法的准确性和精密性良好,可特异性检测Hib PRP.
목적 건립검측b형류감기혈간균(Haemophilus influenzae type b,Hib)다취핵당기핵당순린산염(polyribosylribitol phosphate,PRP)적경쟁매련면역흡부법(competative enzyme-linked immunosorbent assay,cELISA).방법 이Hib PRP-락알(PRP-Ty)위포피항원,대측PRP위경쟁항원,이용방진적정법학정항원여혈청항PRP항체적최괄반응조건,건립cELISA법병험증기준학성화정밀도,병장해법여전통검측법진행비교.결과 최괄포피PRP-Ty농도위1.30 mg/L,혈청항PRP항체희석도위1∶40 000.해법적검측선성범위위6.25～100.00 mg/L,결정계수(R2)위0.99,비내정밀도위3.39％～6.53％,비간정밀도위8.48％.해법대Hib PRP적검측결과여전통화학법일치.결론 건립적Hib다당cELISA법적준학성화정밀성량호,가특이성검측Hib PRP.
Objective To establish competitive ELISA (cELISA) method for determining polyribosylribitol phosphate (PRP) of Haemophilus influenzae type b (Hib).Methods PRP-Ty was used as coating antigen and Hib PRP sample was used as competitive antigen.Optimal reaction conditions were determined between antigens and serum antibodies to Hib PRP by criss-cross serial dilution analysis.The cELISA method was established and its accuracy and precision were validated.The cELISA method was compared with the traditional method.Results Optimal coating concentration of PRP-Ty and antiserum dilution were 1.30 mg/L and 1 ∶ 40 000,respectively.The linear range of the cELISA method was from 6.25 mg/L to 100.00 mg/L,and the coefficient of determination (R2) was 0.99.The precision of intraassay and inter-assay were 3.39％-6.53％ and 8.48％,respectively.The detection results of Hib PRP with the cELISA method were consistent with those with the traditional chemical method.Conclusion The cELISA method has better accuracy and precision,and can be used for detection of Hib polysaccharide.