一种使用地高辛标记探针原位检测巨细胞病毒DNA的杂交免疫细胞化学检测法。

A hybrido-immunocytochemical assay for the in situ detection of cytomegalovirus DNA using digoxigenin-labeled probes.

作者信息

Gentilomi G, Musiani M, Zerbini M, Gallinella G, Gibellini D, La Placa M

机构信息

Institute of Microbiology, University of Bologna, Italy.

出版信息

J Immunol Methods. 1989 Dec 20;125(1-2):177-83. doi: 10.1016/0022-1759(89)90091-4.

DOI:10.1016/0022-1759(89)90091-4

PMID:2558136

Abstract

A non-radioactive hybrido-immunocytochemical assay for the detection of cytomegalovirus (CMV) DNA in infected cells was developed. Two different DNA fragments belonging to the repeated sequences of CMV genome were used to construct the hybridization probe. The probe was constructed by incorporating deoxyuridine triphosphate labeled with digoxigenin. The in situ hybridized CMV DNA probe was immunocytochemically visualized by anti-digoxigenin. Fab fragments labeled with alkaline phosphatase. This procedure permitted the DNA detection, in the nuclei of infected cells fixed at 48 h after infection, of the Towne CMV reference strain and 21 different laboratory-isolated CMV strains. Our assay demonstrated a high specificity, sensitivity and reproducibility.

摘要

开发了一种用于检测感染细胞中巨细胞病毒（CMV）DNA的非放射性杂交免疫细胞化学检测方法。使用属于CMV基因组重复序列的两个不同DNA片段构建杂交探针。该探针通过掺入用地高辛标记的三磷酸脱氧尿苷来构建。原位杂交的CMV DNA探针通过抗地高辛Fab片段用碱性磷酸酶标记进行免疫细胞化学可视化。该方法能够在感染后48小时固定的感染细胞核中检测到Towne CMV参考菌株和21种不同的实验室分离的CMV菌株。我们的检测方法具有高特异性、敏感性和可重复性。