CAJ | 학술논문

目的：应用光激化学发光免疫分析技术（AlphaLISA）建立仙台病毒检测方法。方法通过方阵实验筛选仙台病毒 AlphaLISA 检测的最佳抗原浓度以及供体微珠与受体微珠的最佳受试浓度及比例，对大鼠血清进行测试，确立血清检测浓度；对40份大鼠血清分别使用 AlphaLISA 检测方法和 ELISA 检测方法进行检测，并对检测结果进行比较。结果生物素化标记的仙台病毒多肽类抗原的最佳测试浓度为250 nmol／L，供体微珠与受体微珠的最佳比例为1：1，使用浓度为20μg／mL，在 AlphaLISA 检测方法中大鼠血清测试最佳使用浓度1：10000；共检测40份大鼠血清，ELISA 方法检出阳性7份，阳性率为17．5％，AlphaLISA 方法检出阳性8份，阳性率为20．0％， ELISA 检测阳性的7份血清经 AlphaLISA 方法检测均为阳性，AlphaLISA 检出的另外一份样品经 IFA 方法验证为阳性。结论初步建立了仙台病毒光激化学发光免疫分析技术（AlphaLISA）的检测方法，该方法敏感性堪比经典ELISA 方法，且血清样本的用量减少，无需洗涤等步骤，在方法的简并性及准确性上有一定优势。
목적：응용광격화학발광면역분석기술（AlphaLISA）건립선태병독검측방법。방법통과방진실험사선선태병독 AlphaLISA 검측적최가항원농도이급공체미주여수체미주적최가수시농도급비례，대대서혈청진행측시，학립혈청검측농도；대40빈대서혈청분별사용 AlphaLISA 검측방법화 ELISA 검측방법진행검측，병대검측결과진행비교。결과생물소화표기적선태병독다태류항원적최가측시농도위250 nmol／L，공체미주여수체미주적최가비례위1：1，사용농도위20μg／mL，재 AlphaLISA 검측방법중대서혈청측시최가사용농도1：10000；공검측40빈대서혈청，ELISA 방법검출양성7빈，양성솔위17．5％，AlphaLISA 방법검출양성8빈，양성솔위20．0％， ELISA 검측양성적7빈혈청경 AlphaLISA 방법검측균위양성，AlphaLISA 검출적령외일빈양품경 IFA 방법험증위양성。결론초보건립료선태병독광격화학발광면역분석기술（AlphaLISA）적검측방법，해방법민감성감비경전ELISA 방법，차혈청양본적용량감소，무수세조등보취，재방법적간병성급준학성상유일정우세。
Objective To establish the amplified luminescent proximity homogeneous assay(AlphaLISA) for the detection of Sendai virus.Methods The antigen concentration,serum concentration and the donor beads/acceptor beads ratio used in the AlphaLISA method were optimumized by the phalanx experiments, then the antibodies of Sendai Virus in 40 rat sera were detected by the established AlphaLISA method and ELISA detection method.The results were compared and the differentia between the two methods was confirmed by IFA.Results The optimum concentration of SV bio-peptide in AlphaLISA assay was 250 nmol/L, the best proportion of donor beads/acceptor beads ratio was 1 ∶1, using the concentration of 20 μg/mL and the serum dilution was 1∶10000.7 of the 40 rat sera were detected SV positive by ELISA, the positive rate was 17.5%, 8 of the 40 rat sera were determined SV positive by AlphaLISA, and the positive rate was 20.0%, the AlphaLISA positive serum was confirmed by IFA.Conclusions We preliminary established the Amplified Luminescent Proximity Homogeneous Assay(AlphaLISA) for the detection of Sendai virus.The sensitivity of the method is comparable to classical ELISA method, but this method use less serum samples and without washing steps.The method has some advantages in degeneracy and accuracy.