Detection of the JAK2 V617F mutation by LightCycler PCR and probe dissociation analysis
Detection of the JAK2 V617F mutation by LightCycler PCR and probe dissociation analysis
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DOI：
10.2353/jmoldx.2006.050130
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发表时间：
2006-07-01
影响因子：
4.1
通讯作者：
Zehnder, James L.
中科院分区：
文献类型：
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作者：
Lay, Marla;Mariappan, Rajan;Zehnder, James L.
JAK2基因（酪氨酸激酶家族的一员）的一个点突变最近被发现并被证明与几种骨髓增生性疾病有关。几项研究发现了相同的JAK2点突变（1849G > T），导致密码子617 （V617F）上的缬氨酸被苯丙氨酸取代。我们利用LightCycler平台开发了一种简单、灵敏的方法，通过聚合酶链反应和探针解离分析来检测该突变，并将该方法与现有的限制性片段长度多态性、直接测序和扩增难解突变系统方法进行了比较。我们发现LightCycler方法比限制性片段长度多态性和测序方法具有速度、可靠性和更直接的解释优势。
A point mutation in the JAK2 gene, a member of the tyrosine kinase family, was recently identified and shown to be associated with several myeloproliferative disorders. Several studies identified the same JAK2 point mutation (1849G > T), resulting in the substitution of a valine to phenylalanine at codon 617 (V617F). We developed a simple and sensitive method to detect this mutation via polymerase chain reaction and probe dissociation analysis using the LightCycler platform, and we compared this method to existing restriction fragment-length polymorphism, direct sequencing, and amplification refractory mutation system methods. We found that the LightCycler method offered advantages of speed, reliability, and more straightforward interpretation over the restriction fragment-length polymorphism and sequencing approaches.