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基于聚合酶链反应的微卫星多态性在新鲜及存档肿瘤组织杂合性缺失检测中的应用

PCR-based microsatellite polymorphisms in the detection of loss of heterozygosity in fresh and archival tumour tissue.

作者信息

Gruis N A, Abeln E C, Bardoel A F, Devilee P, Frants R R, Cornelisse C J

机构信息

MGC-Department of Human Genetics, Leiden University, The Netherlands.

出版信息

Br J Cancer. 1993 Aug;68(2):308-13. doi: 10.1038/bjc.1993.333.

DOI:10.1038/bjc.1993.333
PMID:8102243
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1968571/
Abstract

PCR-based microsatellite polymorphisms have proved their power in genetic linkage analysis and other identification methods, due to their high information content and even distribution over the chromosomes. In the present study we applied microsatellite polymorphisms to detect loss of heterozygosity in fresh (snap-frozen) and in archival ovarian tumour tissue. Clear allele losses were found in fresh and paraffin embedded tumour samples. Conventional Southern analysis of flanking markers on the same tumour DNA samples confirmed the observed losses detected by microsatellite polymorphisms. Titration experiments suggest that loss of heterozygosity remains detectable in tumour samples despite 60% contamination with normal DNA. This technique provides a fast and reproducible alternative to conventional Southern blotting in the detection of loss of heterozygosity, with the crucial additional advantages of minimal sample requirements, making archival material available for genetic investigation.

摘要

基于聚合酶链反应（PCR）的微卫星多态性因其高信息含量以及在染色体上的均匀分布，已在遗传连锁分析和其他鉴定方法中展现出强大作用。在本研究中，我们应用微卫星多态性来检测新鲜（速冻）和存档的卵巢肿瘤组织中的杂合性缺失。在新鲜和石蜡包埋的肿瘤样本中均发现了明确的等位基因缺失。对同一肿瘤DNA样本侧翼标记进行的传统Southern分析证实了微卫星多态性检测到的观察到的缺失。滴定实验表明，尽管肿瘤样本被正常DNA污染了60%，但仍可检测到杂合性缺失。该技术在检测杂合性缺失方面为传统Southern印迹法提供了一种快速且可重复的替代方法，具有关键的额外优势，即样本需求量极少，使存档材料可用于基因研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/bff5d351a5f3/brjcancer00198-0098-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/101b3e5ec42e/brjcancer00198-0095-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/c671a5cc21c2/brjcancer00198-0096-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/38c450357b5c/brjcancer00198-0097-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/bff5d351a5f3/brjcancer00198-0098-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/101b3e5ec42e/brjcancer00198-0095-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/c671a5cc21c2/brjcancer00198-0096-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/38c450357b5c/brjcancer00198-0097-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87d1/1968571/bff5d351a5f3/brjcancer00198-0098-a.jpg

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