Point-of-care rapid detection of Vibrio parahaemolyticus in seafood using loop-mediated isothermal amplification and graphene-based screen-printed electrochemical sensor

被引:97
|
作者
Kampeera, Jantana [1 ]
Pasakon, Patiya [2 ]
Karuwan, Chanpen [2 ]
Arunrut, Narong [1 ]
Sappat, Assawapong [2 ]
Sirithammajak, Sarawut [1 ]
Dechokiattawan, Nipaphorn [3 ]
Sumranwanich, Thitinun [3 ]
Chaivisuthangkura, Parin [4 ]
Ounjai, Puey [3 ,5 ]
Chankhamhaengdecha, Surang [3 ]
Wisitsoraat, Anurat [2 ]
Tuantranont, Adisorn [2 ]
Kiatpathomchai, Wansika [1 ]
机构
[1] NSTDA, Natl Ctr Genet Engn & Biotechnol BIOTEC, Bioengn & Sensing Technol Res Team, Pathum Thani 12120, Thailand
[2] NSTDA, Graphene & Printed Elect Dual Use Applicat Res Di, Pathum Thani 12120, Thailand
[3] Mahidol Univ, Dept Biol, Fac Sci, Bangkok 10400, Thailand
[4] Srinakharinwirot Univ, Fac Sci, Dept Biol, Bangkok 10110, Thailand
[5] Minist Educ, Ctr Excellence Environm Hlth & Toxicol, Bangkok 10400, Thailand
来源
关键词
Vibrio parahaemolyticus; LAMP; Screen-printed graphene electrode; Point-of-care; Detection; Seafood; TAURA SYNDROME VIRUS; CARBON ELECTRODE; HIGH-PERFORMANCE; VISUAL DETECTION; DNA BIOSENSOR; FABRICATION; ASSAY; PCR; IDENTIFICATION; BACTERIA;
D O I
10.1016/j.bios.2019.02.060
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Vibrio parahaemolyricus is one of the most important foodbome pathogens that cause various life-threatening diseases in human and animals. Here, we present a rapid detection platform for V. parahaemolyticus by combining loop-mediated isothermal amplification (LAMP) and disposable electrochemical sensors based on screen printed graphene electrodes (SPGEs). The LAMP reactions using primers targeting V. parahaemolyticus toxR gene were optimized at an isothermal temperature of 65 degrees C, providing specific detection of V. parahaemolyticus within 45 min at the detection limit of 0.3 CFU per 25 g of raw seafood. The LAMP amplicons can be effectively detected using unmodified SPGEs, redox active molecules namely Hoechst-33258 and a portable potentiostat. Therefore, the proposed system is particularly suitable as a point-of-care device for on-site detection of foodbome pathogens.
引用
收藏
页码:271 / 278
页数:8
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