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CN101323883B 引物臂TaqMan-MGB探针PCR乙型肝炎病毒检测方法和试剂盒 失效 - 权利终止

引物臂TaqMan-MGB探针PCR乙型肝炎病毒检测方法和试剂盒
摘要：
本发明公开引物臂TaqMan-MGB探针PCR乙型肝炎病毒检测方法和试剂盒，采用在下游引物上加一臂序列，将TaqMan-MGB探针结合在这个臂序列上，结合TaqMan-MGB探针的下游引物和模板结合，在Taq酶的作用下延伸，上游引物结合在第一次循环产物上在Taq酶的作用下延伸，当延伸到第一次循环产物的5′端TaqMan-MGB探针杂交位置时，Taq DNA聚合酶利用其5’-3’外切酶特性沿5’-3’方向切除方向上的障碍物-TaqMan-MGB探针而进一步延长，使TaqMan-MGB探针5’端发光基团[R]游离出来，能量转移现象终止，检测[R]发出的荧光，其荧光强度与PCR产物量成正比。本发明的引物臂TaqMan-MGB探针使两个引物之间的距离尽量缩短，提高检测的敏感性。
摘要（英）：

The invention discloses a detecting method for hepatitis B virus by PCR and using a TaqMan-MGB probe of a primer arm and a kit. A down-stream primer is added with an arm sequence; the TaqMan-MGB probe is combined with the arm sequence; the down-stream primer combined with the TaqMan-MGB probe is combined with a template and extends under the effect of Tag enzyme; an up-stream primer is combined with a product of the first circulation and extends under the effect of Tag enzyme; when the up-stream primer extends to the hybridization position of the TaqMan-MGB probe that is at a 5' end of the product of the first circulation, Tag DNA polymerase excises directional obstacles along a 5' to 3' direction by utilizing characteristics of 5' to 3' excision enzyme; the TaqMan-MGB probe further extends, and luminophore (R) at the 5' end of the TaqMan-MGB probe frees; the phenomenon of energy transfer disappears; the fluorescence emitted by (R) is detected, and fluorescence intensity is in direct proportion to the amount of PCR products. The TaqMan-MGB probe of the primer arm shortens the distance between two primers and promotes detection sensitivity.

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