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使用生物素和地高辛配基标记探针的间期细胞遗传学：III. 提高检测宫颈活检标本和细胞系中HPV的灵敏度和灵活性。

Interphase cytogenetics using biotin and digoxigenin labelled probes: III. Increased sensitivity and flexibility for detecting HPV in cervical biopsy specimens and cell lines.

作者信息

Herrington C S, Graham A K, McGee J O

机构信息

University of Oxford, Nuffield Department of Pathology and Bacteriology, John Radcliffe Hospital, Headington.

出版信息

J Clin Pathol. 1991 Jan;44(1):33-8. doi: 10.1136/jcp.44.1.33.

DOI:10.1136/jcp.44.1.33
PMID:1847709
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC497011/
Abstract

A monoclonal antibody to digoxin enabled sandwich techniques to be used for the detection of hybridised digoxigenin labelled probes in cultured cells and paraffin wax sections. This system has greater flexibility than alkaline phosphatase conjugated polyclonal antidigoxigenin antibody and permits the use of alternative detector enzymes, such as horseradish peroxidase and fluorescence labels. The APAAP detection system that does not require the use of biotin can also be used in situations where endogenous biotin is a problem. The low level of background staining combined with precise substrate deposition of the amplified peroxidase system gives higher sensitivity and resolution. This permits localisation of closely adjacent chromosomal loci in interphase nuclei. The most sensitive peroxidase based digoxigenin detection system visualises two and a half to 12 copies of human papillomavirus (HPV) per nucleus. This system is also suitable for the analysis of low copy number HPV infection of cervical tissues.

摘要

一种地高辛单克隆抗体使得夹心技术可用于检测培养细胞和石蜡切片中杂交的地高辛配体标记探针。该系统比碱性磷酸酶偶联的多克隆抗地高辛配体抗体具有更大的灵活性，并允许使用替代的检测酶，如辣根过氧化物酶和荧光标记物。不需要使用生物素的APAAP检测系统也可用于内源性生物素存在问题的情况。低水平的背景染色与扩增过氧化物酶系统精确的底物沉积相结合，可提供更高的灵敏度和分辨率。这使得在间期核中定位紧密相邻的染色体位点成为可能。基于过氧化物酶的最灵敏的地高辛配体检测系统可检测到每个细胞核中有2.5至12个人乳头瘤病毒（HPV）拷贝。该系统也适用于分析宫颈组织中低拷贝数的HPV感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/1d3e263e9536/jclinpath00403-0045-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/d90ca4e826cf/jclinpath00403-0043-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/240bb82dd2b5/jclinpath00403-0043-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/4afadca55afa/jclinpath00403-0044-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/f294e6fe23e9/jclinpath00403-0044-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/1d3e263e9536/jclinpath00403-0045-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/d90ca4e826cf/jclinpath00403-0043-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/240bb82dd2b5/jclinpath00403-0043-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/4afadca55afa/jclinpath00403-0044-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/f294e6fe23e9/jclinpath00403-0044-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aad9/497011/1d3e263e9536/jclinpath00403-0045-a.jpg

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Detection of chromosome aberrations in the human interphase nucleus by visualization of specific target DNAs with radioactive and non-radioactive in situ hybridization techniques: diagnosis of trisomy 18 with probe L1.84.通过放射性和非放射性原位杂交技术可视化特定靶DNA来检测人类间期核中的染色体畸变：用探针L1.84诊断18三体综合征
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