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通过寡核苷酸溶液杂交和掺入放射性标记的脱氧核苷酸来检测酶促扩增的人类免疫缺陷病毒DNA。

Detection of enzymatically amplified human immunodeficiency virus DNA by oligonucleotide solution hybridization and by incorporation of radiolabeled deoxynucleotides.

作者信息

Carman W F, Williamson C

机构信息

Medical Research Council AIDS Virus Research Unit, University of the Witwatersrand, Johannesburg, South Africa.

出版信息

J Clin Microbiol. 1989 Nov;27(11):2570-3. doi: 10.1128/jcm.27.11.2570-2573.1989.

DOI:10.1128/jcm.27.11.2570-2573.1989
PMID:2808679
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC267078/
Abstract

Two methods are described for the detection of in vitro enzymatically amplified DNA. The first method involves solution hybridization of labeled oligonucleotides to amplified products. Hybridization of primers to a dilution series of known concentration of amplified DNA showed that approximately 5 pg of DNA could be detected by this method. In the second method, radiolabeled deoxynucleotides were incorporated into the elongating DNA chain. Both methods were able to detect amplified products 10 cycles before detection by ethidium bromide staining. Some variations of these techniques are discussed.

摘要

本文描述了两种用于检测体外酶促扩增DNA的方法。第一种方法是将标记的寡核苷酸与扩增产物进行溶液杂交。引物与一系列已知浓度的扩增DNA稀释液杂交结果表明，该方法可检测到约5 pg的DNA。第二种方法是将放射性标记的脱氧核苷酸掺入延伸的DNA链中。两种方法均能在溴化乙锭染色检测前10个循环检测到扩增产物。文中还讨论了这些技术的一些变体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa0d/267078/a79e37373489/jcm00071-0189-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa0d/267078/1ade4c1e666c/jcm00071-0188-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa0d/267078/29a5c4ecdb13/jcm00071-0188-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa0d/267078/a79e37373489/jcm00071-0189-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa0d/267078/1ade4c1e666c/jcm00071-0188-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa0d/267078/29a5c4ecdb13/jcm00071-0188-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa0d/267078/a79e37373489/jcm00071-0189-a.jpg

相似文献

1
Detection of enzymatically amplified human immunodeficiency virus DNA by oligonucleotide solution hybridization and by incorporation of radiolabeled deoxynucleotides.通过寡核苷酸溶液杂交和掺入放射性标记的脱氧核苷酸来检测酶促扩增的人类免疫缺陷病毒DNA。
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本文引用的文献

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An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A.一种通过分析扩增的DNA序列进行遗传性疾病产前诊断的改进方法。应用于甲型血友病。
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Direct cloning and sequence analysis of enzymatically amplified genomic sequences.酶促扩增基因组序列的直接克隆与序列分析
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DNA typing from single hairs.单根毛发的DNA分型
Nature. 1988 Apr 7;332(6164):543-6. doi: 10.1038/332543a0.
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Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.用于镰状细胞贫血诊断的β-珠蛋白基因组序列的酶促扩增及限制性酶切位点分析。
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Identification of HIV-infected seronegative individuals by a direct diagnostic test based on hybridisation to amplified viral DNA.通过基于与扩增病毒DNA杂交的直接诊断测试来鉴定HIV感染的血清阴性个体。
Lancet. 1988 Aug 20;2(8608):418-21. doi: 10.1016/s0140-6736(88)90412-6.
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An assessment of optimal conditions for amplification of HIV cDNA using Thermus aquaticus polymerase.使用嗜热栖热菌聚合酶扩增HIV cDNA的最佳条件评估。
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Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.使用热稳定DNA聚合酶进行引物引导的DNA酶促扩增。
Science. 1988 Jan 29;239(4839):487-91. doi: 10.1126/science.2448875.
10
Identification of human immunodeficiency virus sequences by using in vitro enzymatic amplification and oligomer cleavage detection.利用体外酶促扩增和寡聚物裂解检测鉴定人类免疫缺陷病毒序列。
J Virol. 1987 May;61(5):1690-4. doi: 10.1128/JVI.61.5.1690-1694.1987.