Cubie H A, Leslie E E, Smith S, O'Neill H J, Hart H, Cohen B J, Inglis J M
Regional Virus Laboratory, City Hospital, Edinburgh.
J Clin Pathol. 1993 Sep;46(9):840-5. doi: 10.1136/jcp.46.9.840.
AIMS--To compare the sensitivity, specificity, and practicality of recombinant proteins in serological tests for the detection of human parvovirus B19 IgG and IgM. METHODS--Indirect enzyme linked immunosorbent assays using B19 structural proteins expressed in Escherichia coli were developed for the detection of B19 specific IgG and IgM (rELISA-G and rELISA-M). Cells infected with baculovirus expressing B19 structural proteins were also used in an indirect immunofluorescence assay for IgG and IgM antibodies (IFA-G and IFA-M). Antibody capture radioimmunoassays for IgG and IgM (GACRIA and MACRIA) were used as comparative assays. RESULTS--Twenty nine pools of intravenous immunoglobulin were clearly positive for B19 IgG by rELISA-G and contained low IgG titres by GACRIA. From 113 samples tested by all methods, sensitivities of 92% (77/84) and 97% (68/70) were obtained for ELISA and immunofluorescence, respectively, when compared with GACRIA. One hundred and sixteen samples from patients presenting with rash or arthralgia were compared by MACRIA, rELISA-M, and IFA-M. Sensitivities of both recombinant tests were more than 95%. Despite pretreatment to remove IgG or rheumatoid factor, false positive results were a problem in the rELISA-M but were not seen with the IFA-M. CONCLUSIONS--The limited supply of native antigen has severely restricted the wide application of serology for parvovirus B19. The use of recombinant antigens permitted the introduction of local screening tests which had many advantages, including quicker results and relief of the burden on the Reference Laboratory. The use of rELISA-M for sensitivity and IFA-M for specificity and confirmation proved a useful and practical combination for diagnosis of recent infection with B19, and rELISA-G allowed the immune response to be determined in selected populations.
目的——比较重组蛋白在血清学检测中用于检测人细小病毒B19 IgG和IgM的敏感性、特异性和实用性。方法——开发了使用在大肠杆菌中表达的B19结构蛋白的间接酶联免疫吸附测定法，用于检测B19特异性IgG和IgM（rELISA-G和rELISA-M）。用表达B19结构蛋白的杆状病毒感染的细胞也用于IgG和IgM抗体的间接免疫荧光测定（IFA-G和IFA-M）。IgG和IgM的抗体捕获放射免疫测定（GACRIA和MACRIA）用作比较测定法。结果——通过rELISA-G检测，29组静脉注射免疫球蛋白的B19 IgG呈明显阳性，而通过GACRIA检测其IgG滴度较低。与GACRIA相比，在所有方法检测的113个样本中，ELISA和免疫荧光法的敏感性分别为92%（77/84）和97%（68/70）。通过MACRIA、rELISA-M和IFA-M对116例出现皮疹或关节痛的患者样本进行了比较。两种重组检测方法的敏感性均超过95%。尽管进行了预处理以去除IgG或类风湿因子，但rELISA-M中仍存在假阳性结果问题，而IFA-M中未出现此类问题。结论——天然抗原的供应有限严重限制了细小病毒B19血清学检测的广泛应用。重组抗原的使用使得能够开展局部筛查检测，这些检测具有许多优点，包括结果更快以及减轻参考实验室的负担。使用rELISA-M检测敏感性，IFA-M检测特异性并进行确认，被证明是诊断近期B19感染的一种有用且实用的组合，并且rELISA-G可用于确定特定人群的免疫反应。
