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微量中和试验作为选择用于检测人巨细胞病毒IgG抗体的酶联免疫吸附测定临界值的参考方法。

Microneutralization as a reference method for selection of the cut-off of an enzyme-linked immunosorbent assay for detection of IgG antibody to human cytomegalovirus.

作者信息

Gerna G, Revello M G, Palla M, Percivalle E, Torsellini M

机构信息

Virus Laboratory, Institute of Infectious Diseases, University of Pavia, Italy.

出版信息

Microbiologica. 1992 Apr;15(2):177-81.

PMID:1318488
Abstract

In view of developing an enzyme-linked immunosorbent assay (ELISA) for the determination of IgG antibody to human cytomegalovirus, a rapid microneutralization (Nt) assay was used to test five positive standard sera containing increasing amounts of specific antibody and a negative standard serum. The standard serum containing the minimal amount of detectable Nt antibody was selected as a cut-off standard for the ELISA test. Following preliminary testing on previously characterized sera which gave expected results, the ELISA assay was tested in the field on 992 sera from blood donors. In parallel, sera were tested by Nt and complement fixation (CF). ELISA detected 82 negative and 910 positive sera. Nt gave concordant result except for two ELISA-negative sera, which showed Nt antibody titers of 1:10. The absorbance value of these two sera was just below that of the cut-off. Thus, for ELISA, the sensitivity was 99.8% (910/912) and specificity 100% (80/80). CF gave results concordant with ELISA and Nt, except for 23 sera (2 ELISA- and Nt-negative, and 21 ELISA- and Nt-positive) showing anticomplementary activity. Quantitation of specific ELISA antibody was achieved by interpolation from a calibation curve. Nt appears to be the reference test to establish the ELISA cut-off.

摘要

为开发一种用于检测人巨细胞病毒IgG抗体的酶联免疫吸附测定(ELISA),采用快速微量中和(Nt)试验检测了五种含有不同量特异性抗体的阳性标准血清和一种阴性标准血清。选择含有最低可检测Nt抗体量的标准血清作为ELISA试验的临界标准。在对先前已鉴定血清进行初步检测并获得预期结果后,对992份献血者血清进行了现场ELISA检测。同时,采用Nt试验和补体结合(CF)试验对血清进行检测。ELISA检测出82份阴性血清和910份阳性血清。除两份ELISA阴性血清外,Nt试验结果与之相符,这两份血清的Nt抗体效价为1:10。这两份血清的吸光度值略低于临界值。因此,ELISA的灵敏度为99.8%(910/912),特异性为100%(80/80)。除23份显示抗补体活性的血清(2份ELISA和Nt均为阴性,21份ELISA和Nt均为阳性)外,CF试验结果与ELISA和Nt相符。通过从校准曲线插值实现ELISA特异性抗体的定量。Nt似乎是确定ELISA临界值的参考试验。

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