Schirm J, Luijt D S, Pastoor G W, Mandema J M, Schröder F P
Regional Public Health Laboratory, University Hospital, Groningen, The Netherlands.
J Med Virol. 1992 Oct;38(2):147-51. doi: 10.1002/jmv.1890380214.
Eleven hundred and thirty-three clinical specimens submitted to the laboratory for diagnosis of respiratory virus infections were tested by direct immunofluorescence (DIF) for respiratory syncytial virus (RSV), by shell vial culture, and by conventional cell culture. The shell vial cultures were stained with 8 different monoclonal antibodies both 1 day and 3-7 days after inoculation. In order to limit the cost and the workload, mixtures of monoclonal antibodies were used. Coverslips with HEp-2 cells were incubated with a mixture of FITC-labeled monoclonal antibody to RSV and nonlabeled monoclonal antibody to adenovirus. When no RSV positive IF staining was observed after the first incubation step, the same coverslip was incubated once more with FITC-labeled anti-mouse antibody. A positive reaction at this stage indicated the presence of adenovirus. Similarly, cultures of tertiary monkey kidney cells were investigated with a mixture of two FITC-labeled monoclonals to the influenza viruses A and B and three nonlabeled monoclonals to the parainfluenza viruses 1, 2 and 3. If influenza virus or parainfluenza virus was detected, the exact type was determined by staining different parts of a duplicate coverslip. Shell vial cultures for cytomegalovirus (CMV) were always performed separately on human embryonic lung fibroblasts. Using this approach, we detected RSV (n = 248), CMV (n = 42), parainfluenza virus (n = 31), influenza virus (n = 28), and adenovirus (n = 6), in most cases after only one day of culture. For RSV, the sensitivity of the shell vial method was too low (74%) to allow omission of DIF (sensitivity 95%).(ABSTRACT TRUNCATED AT 250 WORDS)
1133份提交至实验室用于诊断呼吸道病毒感染的临床标本,通过直接免疫荧光法(DIF)检测呼吸道合胞病毒(RSV),采用空斑小瓶培养法及传统细胞培养法。空斑小瓶培养物在接种后1天及3 - 7天用8种不同的单克隆抗体进行染色。为限制成本和工作量,使用了单克隆抗体混合物。将带有HEp - 2细胞的盖玻片与异硫氰酸荧光素(FITC)标记的抗RSV单克隆抗体和未标记的抗腺病毒单克隆抗体混合物孵育。若在第一步孵育后未观察到RSV阳性免疫荧光染色,则将同一块盖玻片再次与FITC标记的抗小鼠抗体孵育。此阶段的阳性反应表明存在腺病毒。同样,用两种FITC标记的针对甲型和乙型流感病毒的单克隆抗体以及三种未标记的针对副流感病毒1、2和3的单克隆抗体混合物研究三级猴肾细胞培养物。若检测到流感病毒或副流感病毒,则通过对重复盖玻片的不同部位进行染色来确定确切类型。巨细胞病毒(CMV)的空斑小瓶培养始终在人胚肺成纤维细胞上单独进行。采用这种方法,我们检测到了RSV(n = 248)、CMV(n = 42)、副流感病毒(n = 31)、流感病毒(n = 28)和腺病毒(n = 6),大多数情况下仅培养1天后即可检测到。对于RSV,空斑小瓶法的灵敏度过低(74%),无法省略DIF法(灵敏度95%)。(摘要截断于250字)