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发明公开
CN105441573A 用于检测人类JAK2基因V617F突变的引物、探针及试剂盒 无效 - 驳回

用于检测人类JAK2基因V617F突变的引物、探针及试剂盒
摘要：
本发明公开了用于检测人类JAK2基因V617F突变的引物、探针及试剂盒，其中引物及探针包括突变上游引物JW1-F，与JAK2基因保守序列结合；突变下游ARMS引物JM1-R，与V617F(1849G>T)位点突变序列特异性结合，选择性扩增突变序列；5’端标记有FAM信号3’端标记MGB的检测探针JW1-P，能与扩增片段结合；以及5’端双脱氧修饰3’端MGB标记的阻断探针JW1-B，能与V617F位点野生型序列特异性结合，抑制野生型非特异扩增。本发明的引物及探针特异性高、灵敏度好，检测能力高达1％。由其制备的试剂盒检测结果准确易读，操作简单快速，适用范围广。
摘要（英）：

The invention discloses a primer, a probe and a kit for detecting a mutation of a human JAK2 gene V617F. The primer comprises a mutation forward primer JW1-F and a mutation reverse ARMS primer JM1-R, and the probe comprises a detection probe JW1-P and a blocking probe JW1-B, wherein the mutation forward primer JW1-F is combined with a JAK2 gene conserved sequence; the mutation reverse ARMS primer JM1-R is specifically combined with a V617F (1849G>T) site mutation sequence to selectively amplify the mutation sequence; a 5minute end of the detection probe JW1-P is marked with an FAM (carboxy fluorescein) signal; a 3minute end of the detection probe is marked with an MGB (Minor Groove Binder); the detection probe JW1-P can be combined with an amplified fragment; a 5minute end of the blocking probe JW1-B is double-deoxidized and modified; a 3minute end of the blocking probe JW1-B is marked with the MGB; the blocking probe JW1-B can be specifically combined with a V617 site wild-type sequence to inhibit wild-type nonspecific amplification. The primer and the probe which are disclosed by the invention are high in specificity and good in sensitivity, and have high detectability of 1 percent. The kit prepared from the primer and the probe is accurate in detection result and easy in reading of the detection result, is simple and rapid to operate, and is wide in application range.

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