A LAMP-based ratiometric electrochemical sensing for ultrasensitive detection of Group B Streptococci with improved stability and accuracy

被引:24
作者
Fu, Yixin [1 ]
Zhou, Xiaoyan [3 ]
Duan, Xiaolei [2 ]
Liu, Changjin [2 ]
Huang, Jian [2 ]
Zhang, Tao [2 ]
Ding, Shijia [4 ]
Min, Xun [2 ]
机构
[1] Zunyi Med Univ, Dept Blood Transfus, Affiliated Hosp, Zunyi 563003, Guizhou, Peoples R China
[2] Zunyi Med Univ, Dept Lab Med, Affiliated Hosp, Zunyi 563003, Guizhou, Peoples R China
[3] Qingdao Univ, Affiliated Hosp Med Coll, Dept Clin Lab, Qingdao 266101, Peoples R China
[4] Chongqing Med Univ, Coll Lab Med, Key Lab Clin Lab Diagnost, Minist Educ, Chongqing 400016, Peoples R China
基金
中国国家自然科学基金;
关键词
Electrochemical; Ratiometry; AuNPs@MoS2; Loop-mediated isothermal amplification; Ferrocene; Methylene blue; MEDIATED ISOTHERMAL AMPLIFICATION; SENSITIVE DETECTION; METHYLENE-BLUE; DNA; BIOSENSOR; GRAPHENE; STRATEGY; SEQUENCE; BINDING;
D O I
10.1016/j.snb.2020.128502
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A LAMP ratiometric electrochemical sensor (LAMP E-sensor) for the highly sensitive detection of Group B Streptococci (GBS) was developed based on recycling amplification via LAMP and a dual-signal electrochemical readout. In this LAMP E-sensor, a thiolated beta-cyclodextrin (SH-beta-CD) was immobilized onto the AuNPs@MoS2-modified electrode surface via Au-S bonding. By using a set of four specially designed primers to amplify the target gene (GBS) in less than 1 h, abundant products modified with ferrocene (Fc) were obtained. These Fcmodified products could connect beta-CD via the host-guest interaction between Fc and beta-CD. Additionally, methylene blue (MB) was embed in the duplex of the LAMP products, producing a strong electrochemical signal. Using this MB as a reporter and Fc as an internal control, the LAMP E-sensor showed a broad detection range from 1 fg mu L-1 to 100 pg mu L-1 and a lower detection limit of 0.23 fg mu L-1. Moreover, testing data from clinical samples showed a high correlation between the developed LAMP E-sensor and RT-PCR results. In conclusion, with improved sensitivity and good reproducibility, this LAMP E-sensor may be a promising alternative for GBS detection in the field of clinical pathogen-based molecular diagnostics.
引用
收藏
页数:8
相关论文
共 40 条
[1]   Pathogenic Bacteria Detection Using RNA-Based Loop-Mediated Isothermal-Amplification-Assisted Nucleic Acid Amplification via Droplet Microfluidics [J].
Azizi, Morteza ;
Zaferani, Meisam ;
Cheong, Soon Hon ;
Abbaspourrad, Alireza .
ACS SENSORS, 2019, 4 (04) :841-848
[2]  
Bannister Stephanie A, 2018, High Throughput, V7, DOI 10.3390/ht7040032
[3]   Genomagnetic LAMP-based electrochemical test for determination of high-risk HPV16 and HPV18 in clinical samples [J].
Bartosik, Martin ;
Jirakova, Ludmila ;
Anton, Milan ;
Vojtesek, Borivoj ;
Hrstka, Roman .
ANALYTICA CHIMICA ACTA, 2018, 1042 :37-43
[4]   Analytical sensitivity and specificity of a loop-mediated isothermal amplification (LAMP) kit prototype for detection of Trypanosoma cruzi DNA in human blood samples [J].
Besuschio, Susana A. ;
Llano Murcia, Monica ;
Benatar, Alejandro F. ;
Monnerat, Severine ;
Cruz, Israel ;
Picado, Albert ;
de los Angeles Curto, Maria ;
Kubota, Yutaka ;
Wehrendt, Diana P. ;
Pavia, Paula ;
Mori, Yasuyoshi ;
Puerta, Concepcion ;
Ndung'u, Joseph M. ;
Schijman, Alejandro G. .
PLOS NEGLECTED TROPICAL DISEASES, 2017, 11 (07)
[5]   Isothermal DNA amplification with functionalized graphene and nanoparticle assisted electroanalysis for rapid detection of Johne's disease [J].
Chand, Rohit ;
Wang, Yi Lan ;
Kelton, David ;
Neethirajan, Suresh .
SENSORS AND ACTUATORS B-CHEMICAL, 2018, 261 :31-37
[6]  
Chen Hong, 2019, CATALYSTS, V9
[7]   Ratiometric Electrochemical Sensor for Effective and Reliable Detection of Ascorbic Acid in Living Brains [J].
Cheng, Hanjun ;
Wang, Xiaoyu ;
Wei, Hui .
ANALYTICAL CHEMISTRY, 2015, 87 (17) :8889-8895
[8]   A reusable ratiometric electrochemical biosensor on the basis of the binding of methylene blue to DNA with alternating AT base sequence for sensitive detection of adenosine [J].
Cui, Lin ;
Lu, Mengfei ;
Li, Ying ;
Tang, Bo ;
Zhang, Chun-yang .
BIOSENSORS & BIOELECTRONICS, 2018, 102 :87-93
[9]   MoS2 nanosheets as an effective fluorescence quencher for DNA methyltransferase activity detection [J].
Deng, Huimin ;
Yang, Xinjian ;
Gao, Zhiqiang .
ANALYST, 2015, 140 (09) :3210-3215
[10]   Reagentless, Ratiometric Electrochemical DNA Sensors with Improved Robustness and Reproducibility [J].
Du, Yan ;
Lim, Byung Joon ;
Li, Bingling ;
Jiang, Yu Sherry ;
Sessler, Jonathan L. ;
Ellington, Andrew D. .
ANALYTICAL CHEMISTRY, 2014, 86 (15) :8010-8016