CAJ | 학술논문

目的参考美国病理学家协会(CAP)相关要求,验证本实验室自建的JAK2 V617F突变检测项目的各项性能参数.方法方法学建立.收集2010年4月至12月复旦大学附属华山医院已确诊的57例BCR/ABL融合基因阴性骨髓增殖性肿瘤患者的外周血和骨髓液标本.采用荧光PCR结合熔解曲线法建立JAK2 V617F突变方法检测标本.在准确度、重复性、分析灵敏度、干扰物质的影响等各方面对检测方法的性能进行评估.准确性使用Kappa检验评估,重复性使用符合率,分析灵敏度使用配对t检验进行统计分析.结果荧光PCR结合熔解曲线方法(试验方法)与参考方法(测序法)相比有非常好的一致性(Kappa=0.89)；重复性符合率为100％；血脂(＜24 mmol/L)和黄疸(＜450 μmol/L)对本试验无明显干扰；最低检测突变率为5％.结论基于荧光PCR结合熔解温度反应方法检测JAK2 V617F突变可应用于临床检测.
목적 삼고미국병이학가협회(CAP)상관요구,험증본실험실자건적JAK2 V617F돌변검측항목적각항성능삼수.방법 방법학건립.수집2010년4월지12월복단대학부속화산의원이학진적57례BCR/ABL융합기인음성골수증식성종류환자적외주혈화골수액표본.채용형광PCR결합용해곡선법건립JAK2 V617F돌변방법검측표본.재준학도、중복성、분석령민도、간우물질적영향등각방면대검측방법적성능진행평고.준학성사용Kappa검험평고,중복성사용부합솔,분석령민도사용배대t검험진행통계분석.결과 형광PCR결합용해곡선방법(시험방법)여삼고방법(측서법)상비유비상호적일치성(Kappa=0.89)；중복성부합솔위100％；혈지(＜24 mmol/L)화황달(＜450 μmol/L)대본시험무명현간우；최저검측돌변솔위5％.결론 기우형광PCR결합용해온도반응방법검측JAK2 V617F돌변가응용우림상검측.
Objective According to the requirements of the College of American Pathologists (CAP),to validate the laboratory developed test-JAK2 V617F mutation.Methods This is a clinical laboratory diagnosis study.Fifty-seven peripheral blood and bone marrow samples were collected from the BCR/ABL-negative myeloproliferative neoplasms (MPN) patients in Huashan Hospital from Apr 2010 to Dec 2010.JAK2 V617F mutation was detected by fluorescence PCR combined with melting curve analysis.The accuracy,precision,analytical sensitivity,and anti-interference ability of the assay were validated.The Kappa test was used to evaluate accuracy.Coincidence rate was used to evaluate the repeatability.Paired t test was used to evaluate analytical sensitivity.Results There was a close agreement between the reference method(sequencing) and the test method (fluorescence melting curve analysis) (Kappa =0.89).The coincidence rate was 100％.The results of the assay was not affected by lipoprotein (＜ 24 mmol/L) or bilirubin (＜450 μmol/L).The detection limit of the assay was 5％.Conclusion JAK2 V617F mutation assay by fluorescence PCR combined with melting curve analysis could be applied to the clinical laboratory.