Provided are a PNA chip for determining the genotypes of mycobacterial species in which probe PNAs having DNA sequences of SEQ ID NOs: 5-28 are immobilized on a plastic substrate coated with an epoxy group-containing polymer, and a method of determining the genotypes of mycobacterial species using the PNA chip. According to the PNA chip and the method of determining the genotypes of mycobacterial species, the genotypes of mycobacterial species which are causes of tuberculosis and respiratory diseases can be determined within a short term with good sensitivity and specificity. Therefore, detection of the presence of mycobacterial species in test samples, and after confirmation of the presence of mycobacterial species, identification of the mycobacterial species and determination of whether or not the mycobacterial species are tuberculosis-causing mycobacterial species can be achieved promptly and accurately.