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[用地高辛标记的双链DNA探针原位杂交检测自然感染犬瘟热脑炎犬脑组织中的犬瘟热病毒N蛋白RNA]

[Detection of distemper virus N protein RNA in the brain of dogs with spontaneous distemper encephalitis using a digoxigenin-labeled, double-stranded DNA probe for in situ hybridization].

作者信息

Gaedke K, Teifke J P, Hardt M, Alldinger S, Baumgärtner W

机构信息

Institut für Veterinär-Pathologie, Justus-Liebig-Universität in Giessen.

出版信息

Berl Munch Tierarztl Wochenschr. 1995 Feb;108(2):51-4.

PMID:7786277
Abstract

A digoxigenin-labelled dsDNA-probe of 287 basepairs length complementary to the nucleoprotein-gene of canine distemper virus (CDV) was generated by the polymerase-chain-reaction. The dsDNA-probe hybridized specifically with base sequences of 8 different CDV strains, whereas no hybridization was observed with a porpoise and a canine parainfluenza virus and only a weak signal was obtained with measles virus. In formalin-fixed, paraffin-embedded brain sections of 35 immunohistologically CDV antigen positive dogs with spontaneous distemper encephalitis CDV-RNA could be detected in 25 cases by in situ hybridization. The reason for the lack of RNA detection in some immunohistologically positive dogs may be due to the low stability of DNA-RNA-hybrids. Degradation of RNA by RNAses or diffusion out of autolysed cells can not be excluded.

摘要

通过聚合酶链反应生成了一段长度为287个碱基对的、与犬瘟热病毒(CDV)核蛋白基因互补的地高辛标记双链DNA探针。该双链DNA探针与8种不同CDV毒株的碱基序列特异性杂交,而与鼠海豚和犬副流感病毒未观察到杂交,与麻疹病毒仅获得微弱信号。在35只患有自发性瘟热脑炎且免疫组织化学检测CDV抗原呈阳性的犬的福尔马林固定、石蜡包埋脑切片中,通过原位杂交在25例中检测到了CDV-RNA。一些免疫组织化学检测呈阳性的犬未检测到RNA的原因可能是DNA-RNA杂交体的稳定性较低。不能排除RNA酶对RNA的降解或其从自溶细胞中扩散出去的情况。

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