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利用互补DNA探针鉴定鼻病毒

Identification of rhinoviruses by cDNA probes.

作者信息

Auvinen P, Ziegler T, Skern T, Kuechler E, Stanway G, Hyypiä T

机构信息

Department of Virology, University of Turku, Finland.

出版信息

J Virol Methods. 1990 Jan;27(1):61-8. doi: 10.1016/0166-0934(90)90146-7.

Abstract

We have used nucleic acid hybridization for the detection and grouping of human rhinoviruses (HRV) according to their genetic relationships. Fifteen rhinovirus reference strains, seventy-one clinical isolates and four enteroviruses were propagated in cell cultures, spotted onto membrane filters and hybridized with radioactively labelled cDNA probes covering different parts of the genomes of HRV-1B, HRV-2, HRV-14, HRV-85 and HRV-89. When the rhinovirus and enterovirus reference strains were tested, the 5' probe of HRV-2 hybridized with thirteen of the fifteen HRV reference strains, with poliovirus type 3 and with ECHO virus 11. The HRV-14 5' probe reacted with eleven HRV reference strains and with all the enteroviruses studied. Sixty-nine of the 71 clinical isolates were recognised by the HRV-2 5' probe, whereas the HRV-14 probe from the same part of the genome hybridized with 54 field isolates. One of the two isolates that remained negative with the HRV-2 5' probe was detected with the HRV-2 probe that derived from the P2 region of the genome, and the other isolate was not detected by any of the probes. Probes from other parts than the 5' end of the genome were generally more specific, and clusters could be formed based on the reactivity of the HRV strains with these probes.

摘要

我们已利用核酸杂交技术,根据人鼻病毒(HRV)的遗传关系对其进行检测和分组。15株鼻病毒参考毒株、71株临床分离株和4株肠道病毒在细胞培养物中增殖,点样于膜滤器上,并与覆盖HRV-1B、HRV-2、HRV-14、HRV-85和HRV-89基因组不同部分的放射性标记cDNA探针杂交。对鼻病毒和肠道病毒参考毒株进行检测时,HRV-2的5'探针与15株HRV参考毒株中的13株、3型脊髓灰质炎病毒和埃可病毒11发生杂交。HRV-14的5'探针与11株HRV参考毒株以及所有研究的肠道病毒发生反应。71株临床分离株中的69株可被HRV-2的5'探针识别,而来自基因组同一部分的HRV-14探针与54株现场分离株杂交。未被HRV-2的5'探针检测到的2株分离株中的1株,被来自基因组P2区域的HRV-2探针检测到,另一株分离株未被任何探针检测到。来自基因组5'端以外其他部分的探针通常更具特异性,可根据HRV毒株与这些探针的反应性形成聚类。

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