Development of an immunoFET biosensor for the detection of biotinylated PCR product.
ImmunoFET (IMFET) biosensor is a simple platform for the detection of biotinylated products of polymerase chain reaction (PCR). Construction of the IMFET biosensor started with adsorption of 1.5 mg/mL of protein A (PA) onto the insulated gate surface of ISFET for 90 min. Next, the immobilized 1/500 dilution of anti-biotin antibody was adsorbed onto the PA layer for 60 min. The IMFET biosensor was subsequently ready for detection of the biotinylated amplicon. The IMFET biosensor showed highly specific binding to the biotinylated PCR product of thephaEgene ofHaloquadratum walsbyiDSM 16854. ThephaEgene is a biomarker of polyhydroxyalkanoate (PHA) producers that contain PHA synthase class III. The lowest amount of DNA template ofH. walsbyiDSM 16854 that the IMFET biosensor could detect was 125 fg. The IMFET biosensor has a lower amount of detection compared with a DNA lateral flow biosensor from our previous study. The degree of linearity of the biosensor signal was influenced by the concentration of the biotinylated amplicon. The IMFET biosensor also has a short response time (approximately 30 times) to detect thephaEamplicon compared to an agarose gel electrophoresis. The IMFET biosensor is a promising tool for the detection of the biotinylated PCR product, and it can be integrated into a micro total analysis system (μTAS).
Department of Biochemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Road, Ratchathewi, Bangkok 10400, Thailand.
Wannaporn
W
Muangsuwan
Department of Biomedical Engineering, Faculty of Engineering, Mahidol University, 999 Phutthamonthon 4 Road, Salaya, Phutthamonthon, Nakhon Pathom 73170, Thailand.
Chamras
C
Promptmas
Thai Microelectronics Center (TMEC), 51/4 Moo 1 Suwintawong Road, Wangtakien, Muang, Chachoengsao 24000, Thailand.
Wutthinan
W
Jeamsaksiri
Thai Microelectronics Center (TMEC), 51/4 Moo 1 Suwintawong Road, Wangtakien, Muang, Chachoengsao 24000, Thailand.
Win
W
Bunjongpru
Thai Microelectronics Center (TMEC), 51/4 Moo 1 Suwintawong Road, Wangtakien, Muang, Chachoengsao 24000, Thailand.
Awirut
A
Srisuwan
Thai Microelectronics Center (TMEC), 51/4 Moo 1 Suwintawong Road, Wangtakien, Muang, Chachoengsao 24000, Thailand.
Charndet
C
Hruanun
Thai Microelectronics Center (TMEC), 51/4 Moo 1 Suwintawong Road, Wangtakien, Muang, Chachoengsao 24000, Thailand.
Amporn
A
Poyai
Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand.
Prapimpun
P
Wongchitrat
Department of Biochemistry, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Road, Ratchathewi, Bangkok 10400, Thailand; Chulabhorn Graduate Institute, 54 Kamphaeng Phet 6, Talat Bang Khen, Lak Si, Bangkok 10210, Thailand.
Montri
M
Yasawong
Biochemistry; Bioengineering; Biotechnology; Engineering; Microbiology