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在微量培养瓶细胞培养系统中通过荧光单克隆抗体染色和原位DNA杂交快速检测巨细胞病毒

Rapid detection of cytomegalovirus by fluorescent monoclonal antibody staining and in situ DNA hybridization in a dram vial cell culture system.

作者信息

Sorbello A F, Elmendorf S L, McSharry J J, Venezia R A, Echols R M

机构信息

Department of Medicine, Albany Medical College, New York 12208.

出版信息

J Clin Microbiol. 1988 Jun;26(6):1111-4. doi: 10.1128/jcm.26.6.1111-1114.1988.

Abstract

By using dram vial cell culture methods, three commercially available tests for cytomegalovirus (CMV) detection were compared: direct fluorescent monoclonal antibody staining for CMV-specific early and late antigens (direct FA), indirect fluorescent monoclonal antibody staining for a CMV-specific early antigen (indirect FA), and in situ DNA hybridization with a biotinylated CMV-specific DNA probe kit (DNA probe). Of those tests, only the indirect FA provided consistent, reliable virus detection within the initial 24 h postinfection for serial 10-fold dilutions of CMV AD169 (laboratory strain) and for three selected urine samples. However, when used prospectively, the indirect FA failed to detect virus within the initial 10 days postinfection in 15 of 78 consecutive specimens that were eventually positive by cell culture. Although the indirect FA was more sensitive than the direct FA or DNA probe, its utility appeared limited to specimens with high CMV concentrations. On the basis of these data, we recommend that indirect FA be reserved as an adjunct to standard cell culture for selected samples in diagnostic hospital laboratories.

摘要

通过使用微量瓶细胞培养方法,比较了三种市售的巨细胞病毒(CMV)检测试验:针对CMV特异性早期和晚期抗原的直接荧光单克隆抗体染色(直接荧光抗体法)、针对CMV特异性早期抗原的间接荧光单克隆抗体染色(间接荧光抗体法)以及使用生物素化的CMV特异性DNA探针试剂盒进行的原位DNA杂交(DNA探针法)。在这些试验中,对于CMV AD169(实验室毒株)的连续10倍稀释液以及三份选定的尿液样本,只有间接荧光抗体法能够在感染后最初24小时内提供一致且可靠的病毒检测结果。然而,在进行前瞻性检测时,在最终经细胞培养呈阳性的78份连续标本中,有15份在感染后最初10天内间接荧光抗体法未能检测到病毒。虽然间接荧光抗体法比直接荧光抗体法或DNA探针法更灵敏,但其应用似乎仅限于CMV浓度较高的标本。基于这些数据,我们建议在诊断医院实验室中,间接荧光抗体法应作为标准细胞培养的辅助方法用于选定样本。

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