A novel linear displacement isothermal amplification with strand displacement probes (LDIA-SD) in a pocket-size device for point-of-care testing of infectious diseases
文献类型： 外文期刊
作者： Gou, Hongchao 1 ; Lin, Qijie 2 ; Shen, Haiyan 1 ; Jia, Kaiyuan 2 ; Liang, Yucen 2 ; Peng, Junhao 2 ; Zhang, Chunhong 1 ; Qu, Xiaoyun 2 ; Li, Yanbin 3 ; Lin, Jianhan 4 ; Zhang, Jianmin 2 ; Liao, Ming 1 ;
作者机构： 1.Inst Anim Hlth, Guangdong Acad Agr Sci, Guangdong Prov Key Lab Livestock Dis Prevent, Maoming Branch,Guangdong Lab Lingnan Modern Agr,ci, Guangzhou 510640, Peoples R China
2.South China Agr Univ, Coll Vet Med, Natl & Reg Joint Engn Lab Medicament Zoonoses Prev, Key Lab Zoonoses Minist Agr,Key Lab Zoonoses Preve, Guangzhou 510642, Peoples R China
3.Univ Arkansas, Dept Biol & Agr Engn, Fayetteville, AR 72701 USA
4.China Agr Univ, Key Lab Agr Informat Acquisit Technol, Minist Agr & Rural Affairs, Beijing 100083, Peoples R China
5.Inst Anim Hlth, Guangdong Acad Agr Sci, Guangzhou 510640, Guangdong, Peoples R China
关键词： Nucleic Acid Amplification; Isothermal amplification; Point -of -care testing (POCT); Linear displacement isothermal amplification; (LDIA); Infectious diseases
期刊名称：SENSORS AND ACTUATORS B-CHEMICAL （ 影响因子：8.4； 五年影响因子：7.2 ）
ISSN：
年卷期： 2023 年 379 卷
页码：
收录情况： SCI
摘要： Nucleic acid amplification is crucial for disease diagnosis, especially lethal infectious diseases such as COVID-19. Compared with PCR, isothermal amplification methods are advantageous for point-of-care testing (POCT). However, complicated primer design limits their application in detecting some short targets or sequences with abnormal GC content. Herein, we developed a novel linear displacement isothermal amplification (LDIA) method using two pairs of conventional primers and Bacillus stearothermophilus (Bst) DNA polymerase, and reactions could be accelerated by adding an extra primer. Pseudorabies virus gE (high GC content) and Salmonella fimW (low GC content) genes were used to evaluate the LDIA assay. Using strand displacement (SD) probes, a LDIA-SD method was developed to realize probe-based specific detection. Additionally, we incorporated a nucleic acid -free extraction step and a pocket-sized device to realize POCT applications of the LDIA-SD method. The LDIA-SD method has advantages including facile primer design, high sensitivity and specificity, and applicability for POCT, especially for amplification of complex sequences and detection of infectious diseases.
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