无内含子基因表达的高灵敏度检测：基于核酸序列扩增（NASBA）技术对信使核糖核酸（mRNA）而非基因组脱氧核糖核酸（gDNA）的扩增
Highly sensitive detection of gene expression of an intronless gene: amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA).
作者信息
Heim A, Grumbach I M, Zeuke S, Top B
机构信息
Institut für Virologie, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
出版信息
Nucleic Acids Res. 1998 May 1;26(9):2250-1. doi: 10.1093/nar/26.9.2250.
Abstract
NASBA is an isothermal nucleic acid amplification reaction that amplifies mRNA in a dsDNA background. Although similar to the sensitive reverse transcription/polymerase chain reaction (RT-PCR) in mRNA detection, NASBA is not prone to give false positive results caused by genomic dsDNA. Therefore, NASBA is unique for sensitive detection of transcription of intronless genes, which preclude strategies such as intron spanning primer pairs to control false positive results in RT-PCR. Using NASBA, mRNA of the intronless human interferon-beta gene was demonstrated with a sensitivity of 10 copies, whereas 100 ng genomic DNA gave a negative result.
摘要
核酸序列依赖性扩增技术（NASBA）是一种等温核酸扩增反应，可在双链DNA背景下扩增信使核糖核酸（mRNA）。尽管在mRNA检测方面与灵敏的逆转录/聚合酶链反应（RT-PCR）相似，但NASBA不易产生由基因组双链DNA导致的假阳性结果。因此，NASBA在灵敏检测无内含子基因的转录方面独具特色，而这排除了诸如使用跨越内含子引物对来控制RT-PCR中假阳性结果的策略。使用NASBA技术，无内含子的人类β-干扰素基因的mRNA检测灵敏度可达10个拷贝，而100纳克基因组DNA检测结果为阴性。
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