【摘要】 目的研究建立时间分辨荧光免疫分析法(TrFIA)检测血清中Ⅳ型胶原(CⅣ),并探讨其临床应用价值。方法以CⅣ多克隆抗体包被板条,Eu3+标记CⅣ单抗,发光增强系统为以β-二酮体为主的增强液。建立CⅣ的夹心TrFIA测定法,并检测127例肝病患者和30例正常对照血清中CⅣ含量。结果CⅣ夹心TrFIA法的灵敏度12．8μg/L;批内和批间平均CV分别为4．54％和8．06％;平均回收率为98．6％。30例正常献血员血清CⅣ含量为(46．06±22．21)μg／L,急性肝炎患者为(47．25±22．58)μg/L,慢性轻度肝炎患者为(129．01±53．68)μg/L,慢性中度肝炎患者为(277．90±92．36)μg/L,慢性重度肝炎患者为(413．90±162．24)μg／L,肝硬化患者为(568．60±210．40)μg／L。除急性肝炎外,均显著高于正常对照组(P<0．01)。结论建立的测定CⅣ的夹心TrFIA法灵敏度和特异性高,重复性好。更多还原

【Abstract】 Objective To establish a time-resolved immunofluorometric assay (TRFIA) to detect seurm CⅣ(collagenⅣ). Methods The antibodies to CⅣwere coated on mircoplate and the europium-labeled monoclonal antibody of CⅣ. The luminescent enhancement system was used as enhancement solution which contained mainly 2-naphthoy trifluoroacetone. we established A sandwich time-resolved fluoroimmunoassay (TRFIA) was established to measure the seurm CⅣin 127 patients with hepatitis and 30 normal controls. Results The sensitivity of assay was 12. 8μg/L. The coefficient of variation for inner-batch and inter-batch were 4. 54% and 8. 06%,respectively. The recovery was 98. 6%. The serum level of CⅣwas 46. 06±22. 21μg/L in normal control,47. 25±22. 58μg/L in acute hepatitis, 129.01±53.68μg/L in mild chronic hepatitis,277. 90±92.36μg/L in moderate chronic hepatitis,413.90±162.24μg/L in serious chronic hepatitis,568. 60±210.40μg/L in liver cirrhosis. As compared to normal control,higher concentrations of CIV (P <0. 01) were observed in chronic hepatitis and liver cirrhosis. Conclusion The established sandwich TRFIA for seurm CⅣmeasurement is a high special and sensitive assay.更多还原