Lan kim tuyen (Anoectochilus setaceus Blume) also known as Anoectochilus roxburghii Wall. ex Lindl belongs to Anoectochilus genus which is generally used in treatment of hypertension, diabetes, heart disease, pneumonia, acute and chronic hepatitis, nephritis and other diseasesas... The identification of this precious medicinal orchid is very difficult due to the narrow of the distribution of wild Anoectochilus aqd the scarce of natural reserves and the missing of flower period, traditional identification methods were usually invalid. Recently, the identification of species using DNA barcode is particularly useful to distinguish species when the observed morphology, growth and development, which are not sufficient basis to identify and distinguish species. In this study, the authors studied the possibility of using two chloroplast DNA barcode markers: psbA-trnH, matK and nuclear marker ITS to identify plant samples collected at Tam Dao National Park and Tumorong, Kontum province. Analysis of nucleotide sequences of these DNA barcode regions matK, psbA-trnH and ITS from these collected samplesshowed that, these samples belong to genus Anoectochilus. Using two markers matK and ITS can be able not only to distinguish these three species Anoectochilus roxburghii, Anoectochilus formosanus and Anoectochilus koshunensis but also other species.