Single-walled carbon nanotubes (SWNTs) have been the promising subject of intense research due to their nanoscale dimensions and outstanding electrical and mechanical properties. Recently, flexible biosensors are rapidly emerging for the next generation of human health technology for detecting serious human species such as glucose, cytochrome, nicotinamide adenine dinucleotide (NADH), insulin, amino acids, and so on. In addition, SWNTs based biosensors are being studied in an effort to explore glucose monitoring in blood. In this research, we demonstrate the glucose oxide modified SWNTs flexible biosensor for D-glucose detection. We have employed fluidic assembly process to assemble and integrate SWNTs on lithographically patterned plastic template. Template guided fluidic assembly method is a relatively simple process compared to transfer printing process because it can be applied to wide range of substrates. Highly ordered SWNTs provide excellent electronic properties and high surface area. For the highly dense SWNTs assembly, the influence of humidity on template guided fluidic assembly of horizontally aligned SWNTs has been studied. To apply the fabricated SWNTs based flexible biosensor for D-glucose detecting, GOD is immobilized on the SWNTs surface using a non-covalent bonding. The GOD-modified SWNTs flexible devices were characterized by electrochemical measurements. The electrical signal was monitored while sequentially increasing glucose solution concentration by applying voltage from 0 to 100mV. The SWNTs based flexible biosensor has high electrochemical sensitivity of detection of glucose. So this flexible device can be used in-vivo application.