1. Establishment of scalable nanoliter digital LAMP technology for the quantitative detection of multiple myeloproliferative neoplasm molecular markers.
- Author
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Cao, Guojun, Li, Jinze, Xing, Zhifang, Zhang, Zhiqi, Zhang, Wei, Li, Chuanyu, Li, Longhui, Guo, Zhen, Li, Shuli, Gao, Xu, Ma, Yanchun, Zhou, Lianqun, and Guan, Ming
- Subjects
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POLYMERASE chain reaction , *TUMORS , *CALRETICULIN , *DIAGNOSIS , *BRUGADA syndrome ,BONE marrow cancer - Abstract
• A scalable nanoliter nano-dLAMP technology was established. • The nano-dLAMP was with lower cost, faster speed and more detection throughputs. • The nano-dLAMP was not limited by the annealing temperature differences. • It could be used not only for sensitive detection of MPN, but also for coronavirus and so on. Myeloproliferative neoplasms (MPNs) are a type of chronic hematological tumor accompanied by bone marrow failure or leukemia. A nanoparticle-assisted digital loop-mediated isothermal amplification (nano-dLAMP) platform was established for the analysis of MPNs in this study. Microarray chips with four physical partitions were fabricated for the simultaneous detection of calreticulin type 1 (CALR-1), calreticulin type 2 (CALR-2), and janus kinase 2 V617F (JAK2 V617F) mutations and an internal reference gene. Polymerase chain reaction (PCR) additives and nanoparticles were used to make the traditional loop mediated isothermal amplification (LAMP) suitable for nanoliter-scale amplification. The results suggested that nanoparticles could improve the amplification performance of nanoliter LAMP. Quantitative detection of the main MPN molecular markers could be performed simultaneously in one four-partition microarray chip within 1 h. The detection sensitivity values for CALR-1, CALR-2, and JAK2 V617F were 0.5 %, 0.1 %, and 0.5 % mutation burden, respectively. The agreement between the developed platform and the commercial Quantstudio 3D was high at 99 % (280/281). This accurate, rapid, multiplex, and inexpensive nano-dLAMP platform could be a promising tool for clinical diagnosis in the future. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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