一种利用人线粒体DNA高变非编码区的限制性片段长度多态性（RFLP）和单链构象多态性（SSCP）分析的快速简便的DNA指纹图谱方法。

A rapid and simple DNA fingerprinting method using RFLP and SSCP analysis of the hypervariable noncoding region of human mitochondrial DNA.

作者信息

Pushnova E A, Akhmedova S N, Shevtsov S P, Schwartz E I

机构信息

Center of Molecular Diagnostics, St. Petersburg Pediatric Medical Institute, Russia.

出版信息

Hum Mutat. 1994;3(3):292-6. doi: 10.1002/humu.1380030318.

DOI:10.1002/humu.1380030318

PMID:7912606

Abstract

A simple method for identification of individuals and maternity testing has been developed. This technique includes PCR amplification of the 199-bp hypervariable portion of the noncoding region of human mtDNA, digestion with RsaI and subsequent SSCP analysis of restriction fragments with DNA silver staining. Using this approach we have analysed the DNA fingerprint patterns of the family members. The fingerprints of maternally related individuals appeared to be identical in three generations, while maternally unrelated members of the family showed differences in their fingerprints, either in SSCP or both RFLP and SSCP patterns. Sequencing data have confirmed the results obtained. Further DNA fingerprinting analysis of 19 unrelated mother-child pairs by means of the method described revealed complete identity of the fingerprint patterns within the pairs. The probability of a random fingerprint match for two maternally unrelated individuals was estimated as 8%.

摘要

已开发出一种用于个体识别和母系检测的简单方法。该技术包括对人线粒体DNA非编码区199bp高变区进行PCR扩增，用RsaI酶切，随后对限制性片段进行单链构象多态性（SSCP）分析并进行DNA银染。利用这种方法，我们分析了家庭成员的DNA指纹图谱。在三代人中，母系相关个体的指纹似乎是相同的，而家族中母系无关的成员在指纹上则表现出差异，无论是在SSCP图谱还是在限制性片段长度多态性（RFLP）和SSCP图谱上。测序数据证实了所得结果。通过所述方法对19对无关的母子进行进一步的DNA指纹分析，结果显示每对内部指纹图谱完全相同。两个母系无关个体随机指纹匹配的概率估计为8%。