公开(公告)号：US5861245A

公开(公告)日：1999-01-19

申请号：US471994

申请日：1995-06-06

申请人： Michael McClelland ,  John Thomas Welsh ,  Joseph A. Sorge

发明人： Michael McClelland ,  John Thomas Welsh ,  Joseph A. Sorge

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/689 ,  C12Q2600/156

摘要： A rapid method for generating a set of discrete DNA amplification products characteristic of a genome as a "fingerprint" comprises the steps of: priming target nucleic acid of a genome with an single-stranded primer to form primed nucleic acid such that a substantial degree of internal-mismatching occurs between the primer and the target nucleic acid; amplifying the primed nucleic acid by performing at least one cycle of polymerase chain reaction amplification; and amplifying the product of step (2) by performing at least about 10 cycles of polymerase chain reaction amplification. The method is known as the arbitrarily primed polymerase chain reaction (AP-PCR) method and is suitable for the identification of bacterial species and strains, including Staphylococcus and Streptococcus species, mammals and plants. The method of the present invention can identify species rapidly, using only a small amount of biological material, and does not require knowledge of the nucleotide sequence or other molecular biology of the nucleic acids of the organisms to be identified. Only one primer sequence is required for amplification and/or identification. The method can also be used to generate detectable polymorphisms for use in genetic mapping of animals and humans.

摘要翻译： 用于产生一组特征为基因组的离散DNA扩增产物作为“指纹”的快速方法包括以下步骤：用单链引物引发基因组的靶核酸以形成引发的核酸，使得大部分 在引物和靶核酸之间发生内部错配; 通过进行聚合酶链反应扩增的至少一个循环来扩增引发的核酸; 并通过进行至少约10个循环的聚合酶链反应扩增来扩增步骤（2）的产物。 该方法被称为任意引物的聚合酶链反应（AP-PCR）方法，适用于鉴定细菌和菌株，包括葡萄球菌和链球菌属，哺乳动物和植物。 本发明的方法可以使用仅少量的生物材料快速鉴定物种，并且不需要知道待鉴定的生物体的核酸序列或其他分子生物学。 扩增和/或鉴定只需要一个引物序列。 该方法还可用于产生用于动物和人的遗传图谱的可检测的多态性。