Establishment of scalable nanoliter digital LAMP technology for the quantitative detection of multiple myeloproliferative neoplasm molecular markers
Establishment of scalable nanoliter digital LAMP technology for the quantitative detection of multiple myeloproliferative neoplasm molecular markers
复制标题
建立可扩展纳升数字LAMP技术定量检测多种骨髓增生性肿瘤分子标志物
DOI：
10.1016/j.snb.2021.130493
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发表时间：
2021-08-02
影响因子：
8.4
通讯作者：
Guan, Ming
中科院分区：
文献类型：
--
作者：
Cao, Guojun;Li, Jinze;Guan, Ming
骨髓增生性肿瘤（MPN）是一种伴有骨髓衰竭或白血病的慢性血液肿瘤。本研究建立了一种纳米粒子辅助数字环介导等温扩增（nano-dLAMP）平台，用于分析MPN。制备了具有4个物理分区的微阵列芯片，用于同时检测钙网蛋白1型（CALR-1）、钙网蛋白2型（CALR-2）和janus激酶2 V617 F（JAK 2 V617 F）突变和内参基因。使用聚合酶链反应（PCR）添加剂和纳米颗粒使传统的环介导等温扩增（LAMP）适合于纳升规模的扩增。结果表明，纳米粒子可以提高纳升LAMP的放大性能。该芯片可在1 h内完成MPN主要分子标记的定量检测。CALR-1、CALR-2和JAK 2 V617 F的检测灵敏度值分别为0.5%、0.1%和0.5%突变负荷。开发的平台和商业Quantstudio 3D之间的一致性高达99%（280/281）。这种准确、快速、多重和廉价的nano-dLAMP平台可能是未来临床诊断的有前途的工具。
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.