A DNA (Deoxyribo Nucleic Acid) detection device and a manufacturing method thereof detects DNA by an electrical method without a separate process for detection by using semiconductor microfabrication techniques. The DNA detection device includes a pair of chambers formed on a semiconductor substrate for accommodating a detection sample, a channel connecting the pair of chambers and a lid covering the pair of chambers. According to the present invention, it is possible to attain a DNA detection device, which can be mass-produced from a silicon substrate by using semiconductor manufacturing technology with improved microfabrication techniques, and a manufacturing method thereof.