Weiwei Liu1, Tingting Hu, Yuming Chen, Xinju Zhang, Xiaoye Gu, Ming Guan. 1. Department of Laboratory Medicine, Shanghai Tenth People's Hospital, Tongji University, 301 Yanchang Road, Shanghai, 200072, People's Republic of China, huashanvivian@126.com.
Abstract
BACKGROUND AND OBJECTIVE: JAK2 V617F mutation is a molecular marker for myeloproliferative neoplasms (MPNs). As there are no China Food and Drug Administration-approved assays for the detection of JAK2 V617F 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 detecting JAK2 V617F using tetra-primer amplification refractory mutation system-polymerase chain reaction (tetra-primer ARMS-PCR) combined with melting-curve analysis. METHODS: A 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. RESULTS: There 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 the JAK2 mutation was 1.25 %. CONCLUSIONS: Tetra-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 the JAK2 V617F mutation.
BACKGROUND AND OBJECTIVE: JAK2 V617F mutation is a molecular marker for myeloproliferative neoplasms (MPNs). As there are no China Food and Drug Administration-approved assays for the detection of JAK2 V617F 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 detecting JAK2 V617F using tetra-primer amplification refractory mutation system-polymerase chain reaction (tetra-primer ARMS-PCR) combined with melting-curve analysis. METHODS: A 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. RESULTS: There 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 the JAK2 mutation was 1.25 %. CONCLUSIONS:Tetra-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 the JAK2 V617F mutation.
Authors: Gurunathan Murugesan; Samer Aboudola; Hadrian Szpurka; Mary Ann Verbic; Jaroslaw P Maciejewski; Raymond R Tubbs; Eric D Hsi Journal: Am J Clin Pathol Date: 2006-04 Impact factor: 2.493
Authors: Ross L Levine; Martha Wadleigh; Jan Cools; Benjamin L Ebert; Gerlinde Wernig; Brian J P Huntly; Titus J Boggon; Iwona Wlodarska; Jennifer J Clark; Sandra Moore; Jennifer Adelsperger; Sumin Koo; Jeffrey C Lee; Stacey Gabriel; Thomas Mercher; Alan D'Andrea; Stefan Fröhling; Konstanze Döhner; Peter Marynen; Peter Vandenberghe; Ruben A Mesa; Ayalew Tefferi; James D Griffin; Michael J Eck; William R Sellers; Matthew Meyerson; Todd R Golub; Stephanie J Lee; D Gary Gilliland Journal: Cancer Cell Date: 2005-04 Impact factor: 31.743
Authors: Amy V Jones; Sebastian Kreil; Katerina Zoi; Katherine Waghorn; Claire Curtis; Lingyan Zhang; Joannah Score; Rachel Seear; Andrew J Chase; Francis H Grand; Helen White; Christine Zoi; Dimitris Loukopoulos; Evangelos Terpos; Elisavet-Christine Vervessou; Beate Schultheis; Michael Emig; Thomas Ernst; Eva Lengfelder; Rüdiger Hehlmann; Andreas Hochhaus; David Oscier; Richard T Silver; Andreas Reiter; Nicholas C P Cross Journal: Blood Date: 2005-05-26 Impact factor: 22.113