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使用非放射性合成寡脱氧核苷酸探针通过斑点印迹杂交试验检测轮状病毒。

Rotavirus detection by dot blot hybridization assay using a non-radioactive synthetic oligodeoxynucleotide probe.

作者信息

Fernández J, Sandino A, Yudelevich A, Avendaño L F, Venegas A, Hinrichsen V, Spencer E

机构信息

Unidad de Virología INTA, Universidad de Chile, Santiago.

出版信息

Epidemiol Infect. 1992 Feb;108(1):175-84. doi: 10.1017/s0950268800049621.

Abstract

A synthetic oligodeoxynucleotide of 40 nucleotides corresponding to nucleotides 33-72 of the gene coding for the viral protein VP7 of rotavirus, was used as a nucleic acid probe to develop a non-radioactive hybridization method for rotavirus detection. The probe was labelled at the 3' end with biotin-7-dATP. The sensitivity and specificity of the dot blot hybridization assay for rotavirus detection was evaluated with 303 stool specimens. The results indicate that the hybridization assay has a higher sensitivity than both PAGE and EIA. Among the rotavirus strains tested 37 different electropherotypes were found. The results suggest that rotavirus diagnosis by dot hybridization using a non-radioactive probe may become routine laboratory procedure because it is simple, highly specific and very sensitive.

摘要

一种与轮状病毒病毒蛋白VP7编码基因的33 - 72位核苷酸相对应的40个核苷酸的合成寡脱氧核苷酸,被用作核酸探针,以开发一种用于轮状病毒检测的非放射性杂交方法。该探针在3'端用生物素 - 7 - dATP进行标记。用303份粪便标本评估了用于轮状病毒检测的斑点印迹杂交试验的灵敏度和特异性。结果表明,该杂交试验比PAGE和EIA具有更高的灵敏度。在所测试的轮状病毒菌株中,发现了37种不同的电泳型。结果表明,使用非放射性探针通过斑点杂交进行轮状病毒诊断可能会成为常规实验室程序,因为它简单、高度特异且非常灵敏。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f3e2/2272188/bcbcb13c1cff/epidinfect00031-0180-a.jpg

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