Development and Validation of a Tetra-Primer Amplification Refractory Mutation System-Polymerase Chain Reaction Combined with Melting Analysis-Assay for Clinical JAK2 V617F Mutation Detection
Development and Validation of a Tetra-Primer Amplification Refractory Mutation System-Polymerase Chain Reaction Combined with Melting Analysis-Assay for Clinical JAK2 V617F Mutation Detection
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用于临床 JAK2 V617F 突变检测的四引物扩增难治性突变系统的开发和验证 - 聚合酶链反应结合熔解分析 - 测定
DOI：
10.1007/s40291-014-0111-6
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发表时间：
2014-07
影响因子：
4
通讯作者：
Guan Ming
中科院分区：
文献类型：
--
作者：
Liu Weiwei;Hu Tingting;Chen Yuming;Zhang Xinju;Gu Xiaoye;Guan Ming
背景与目的JAK 2 V617 F突变是骨髓增生性肿瘤（myeloproliferative neoplasia，MPN）的分子标志物。由于中国尚无国家食品药品监督管理局批准的JAK 2 V617 F突变检测方法，因此在临床实施前验证该方法的分析性能对于临床实验室非常重要。方法采用四引物扩增难治性突变系统-聚合酶链反应（tetra-primer ARMS-PCR）结合熔解曲线分析技术，对复旦大学附属华山医院202例MPN患者的外周血标本和20例骨髓穿刺标本进行JAK 2 V617 F检测。的准确性，精密度，重现性，分析灵敏度，检测下限，分析特异性，干扰物质，耐用性，耐用性，可报告范围和报告的本assay.ResultsThere是一个密切的协议之间的参考方法（测序）和熔解曲线分析（κ= 0.89）。精密度为100%，测定结果不受脂蛋白（<27 mmol/L）或胆红素（<450 µmol/L）的影响。结论四引物扩增突变系统-聚合酶链反应（tetra-primer ARMS-PCR）结合熔解曲线分析可用于JAK 2 V617 F突变的检测。
Background and objectiveJAK2V617F mutation is a molecular marker for myeloproliferative neoplasms (MPNs). As there are no China Food and Drug Administration-approved assays for the detection ofJAK2V617F mutation in China, validation of the analytic performance of this assay is important for the clinical laboratory before its clinical implementation. We have established a method for detectingJAK2V617F using tetra-primer amplification refractory mutation system-polymerase chain reaction (tetra-primer ARMS-PCR) combined with melting-curve analysis.MethodsA total of 202 blood samples and 20 bone marrow aspirates were obtained from MPNs patients at Huashan Hospital, Fudan University. The accuracy, precision, reproducibility, analytical sensitivity, lower limit of detection, analytical specificity, interfering substances, ruggedness, robustness, reportable range and reporting of this assay were validated.ResultsThere was a close agreement between the reference method (sequencing) and melting-curve analysis (κ= 0.89). The precision was 100 % and the results of the assay were unaffected by lipoprotein (<27 mmol/L) or bilirubin (<450 µmol/L). The analytical sensitivity of theJAK2mutation was 1.25 %.ConclusionsTetra-primer amplification refractory mutation system-polymerase chain reaction (tetra-primer ARMS-PCR) combined with melting-curve analysis can be used in the clinical laboratory for detection of theJAK2V617F mutation.
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影响因子：
20.3
作者：
Levine, RL;Loriaux, M;Deininger, MWN
通讯作者：
Deininger, MWN
影响因子：
4.1
作者：
Lay, Marla;Mariappan, Rajan;Zehnder, James L.
通讯作者：
Zehnder, James L.
影响因子：
3.8
作者：
Bracco E;Rosso V;Serra A;Carnuccio F;Gaidano V;Nicoli P;Musto P;Saglio G;Frassoni F;Cilloni D
通讯作者：
Cilloni D
影响因子：
20.3
作者：
Steensma, DP;Dewald, GW;Tefferi, A
通讯作者：
Tefferi, A
影响因子：
2.9
作者：
Baris, Ibrahim;Etlik, Ozdal;Baris, Saniye Tugba
通讯作者：
Baris, Saniye Tugba