The present invention provides facile and accurate molecular diagnosis of disease-specific genes capable of the naked eye detection through amplifying the target genes to selectively block the fluid path in a microfluidic device. Specifically, the present invention includes an isothermal amplification of target genes through a rolling circle amplification, a microfluidic device for detecting pathogen genes, and a detection method using the same. Therefore, the present invention can conveniently detect a single target gene, such as a single pathogen, or at the same time, several target genes, such as several pathogens, without complicated mechanical equipment.