Percivalle E, Zavattoni M, Revello M G, Mariani G, Scaravelli C, Gerna G
Virus Laboratory, University of Pavia, Italy.
Microbiologica. 1989 Jul;12(3):203-13.
Direct fluorescent antibody assay (DFA) using monoclonal antibody and enzyme-linked immunosorbent assay (ELISA) for rapid detection of Respiratory Syncytial Virus (RSV) in nasopharyngeal secretions (NPS) were compared with conventional virus isolation and identification procedures in cell cultures. When 134 NPS were examined from infants and young children with acute respiratory tract infection, 42 (31%) were culture-positive for RSV and 31 of these were detected by the appearance of a typical cytopathic effect and identified by DFA either before or after its appearance, whereas 11 were identified as RSV-positive by DFA performed blindly on HEp-2 cell cultures 5 or 10 days after inoculation. DFA for RSV on NPS smears was positive in 33 (26%) cases, from seven of which RSV was not isolated. The same group of 134 NPS was tested for RSV detection by three commercial ELISA kits. The sensitivities of the three ELISA kits when compared with a combination of culture and DFA results, were comparable (53%, 51%, and 47% for Ortho, Kallested, and Abbott, respectively), whereas specificity was 100% for all three assays. In the group of 26 NPS detected as positive by both virus isolation and DFA, 20-22 (77-85% according to different kits) were found positive for RSV by the three ELISA assays. These data suggest that virus isolation is still critical for diagnosis of a fair number of cases of RSV infection. Of the two rapid techniques, DFA is a valuable complementary method, whereas ELISA still lacks sensitivity. However, both DFA and ELISA were able to detect RSV in 7 of 8 young patients with severe respiratory infection (pneumonia, bronchiolitis), thus permitting diagnosis of RSV infection at least two days before culturing.
采用单克隆抗体的直接荧光抗体检测法（DFA）和酶联免疫吸附测定法（ELISA）用于快速检测鼻咽分泌物（NPS）中的呼吸道合胞病毒（RSV），并与细胞培养中的传统病毒分离和鉴定程序进行了比较。对134例患有急性呼吸道感染的婴幼儿的NPS进行检测时，42例（31%）RSV培养呈阳性，其中31例通过典型细胞病变效应的出现而被检测到，并在其出现之前或之后通过DFA进行鉴定，而11例在接种后5天或10天对HEp-2细胞培养物进行盲法DFA检测时被鉴定为RSV阳性。NPS涂片上RSV的DFA检测在33例（26%）病例中呈阳性，其中7例未分离出RSV。使用三种商用ELISA试剂盒对同一组134例NPS进行RSV检测。与培养和DFA结果相结合相比，三种ELISA试剂盒的灵敏度相当（Ortho、Kallested和Abbott分别为53%、51%和47%），而所有三种检测的特异性均为100%。在病毒分离和DFA均检测为阳性的26例NPS组中，三种ELISA检测发现20 - 22例（根据不同试剂盒为77 - 85%）RSV呈阳性。这些数据表明，病毒分离对于相当数量的RSV感染病例的诊断仍然至关重要。在这两种快速检测技术中，DFA是一种有价值的补充方法，而ELISA仍然缺乏灵敏度。然而，DFA和ELISA均能够在8例患有严重呼吸道感染（肺炎、细支气管炎）的年轻患者中的7例中检测到RSV，从而至少在培养前两天确诊RSV感染。
