Establishment of scalable nanoliter digital LAMP technology for the quantitative detection of multiple myeloproliferative neoplasm molecular markers

被引：5

作者：

Cao, Guojun ^{[1
,2
,3
]}

Li, Jinze ^{[3
]}

Xing, Zhifang ^{[1
]}

Zhang, Zhiqi ^{[3
]}

Zhang, Wei ^{[3
]}

Li, Chuanyu ^{[3
]}

Li, Longhui ^{[3
]}

Guo, Zhen ^{[3
]}

Li, Shuli ^{[3
]}

Gao, Xu ^{[3
]}

Ma, Yanchun ^{[2
]}

Zhou, Lianqun ^{[3
]}

Guan, Ming ^{[1
]}

机构：

[1] Fudan Univ, Huashan Hosp, Shanghai Med Coll, Dept Lab Med, 12 Middle Urumqi Rd, Shanghai 200040, Peoples R China

[2] Fudan Univ, Huashan Hosp North, Shanghai Med Coll, Dept Lab Med, Shanghai 201907, Peoples R China

[3] Chinese Acad Sci, Suzhou Inst Biomed Engn & Technol, CAS Key Lab Biomed Diagnost, 88 Keling Rd, Suzhou 215163, Peoples R China

来源：

SENSORS AND ACTUATORS B-CHEMICAL | 2021年 / 346卷

基金：

国家重点研发计划; 中国博士后科学基金; 中国国家自然科学基金;

关键词：

Myeloproliferative neoplasm (MPN); Rapid detection; Quantitative detection; Nanoparticles; Digital loop-mediated isothermal amplification (dLAMP); BOVINE SERUM-ALBUMIN; ISOTHERMAL AMPLIFICATION; GOLD NANOPARTICLES; DNA; PCR; CHIP; QUANTIFICATION; IMPROVEMENT; INHIBITION; MUTATIONS;

D O I：

10.1016/j.snb.2021.130493

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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.

引用

页数：11