使用cDNA和不对称RNA探针通过原位杂交在VERO细胞和白纹伊蚊C6/36细胞中检测布尼亚病毒杰米斯顿株。

Detection of the bunyavirus Germiston in VERO and Aedes albopictus C6/36 cells by in situ hybridization using cDNA and asymmetric RNA probes.

作者信息

Delord B, Poveda J D, Astier-Gin T, Gerbaud S, Fleury H J

机构信息

Laboratoire de Virologie, Université de Bordeaux II, France.

出版信息

J Virol Methods. 1989 Jun;24(3):253-64. doi: 10.1016/0166-0934(89)90037-2.

DOI:10.1016/0166-0934(89)90037-2

PMID:2760165

Abstract

Using Germiston virus infected vertebrate (VERO) and invertebrate (Aedes albopictus C6/36) cells, paraformaldehyde-glutaraldehyde fixative allowed the best preservation of cellular morphology and the highest hybridization signals with cDNA and asymmetric RNA probes against the viral S segment. Asymmetric RNA probes always gave higher sensitivity and better specificity of in situ hybridization than the nick-translated symmetric DNA probe in both vertebrate and invertebrate cells. The study of Aedes albopictus C6/36 cells persistently infected with Germiston virus showed that only a small number of cells contained the S segment, and that the replication and transcription of the S segment took place in the cytoplasm of acutely and persistently infected cells.

摘要

使用杰米斯顿病毒感染的脊椎动物（VERO）和无脊椎动物（白纹伊蚊C6/36）细胞，多聚甲醛-戊二醛固定剂能最佳地保存细胞形态，并与针对病毒S基因片段的cDNA和不对称RNA探针产生最高的杂交信号。在脊椎动物和无脊椎动物细胞中，不对称RNA探针原位杂交的灵敏度总是高于缺口平移对称DNA探针，特异性也更好。对持续感染杰米斯顿病毒的白纹伊蚊C6/36细胞的研究表明，只有少数细胞含有S基因片段，且S基因片段的复制和转录发生在急性和持续感染细胞的细胞质中。