This study aims to develop a benthic macroinvertebrate multimetric index for assessing the ecological quality of streams in Northeastern Thailand. ANOSIM indicated that the benthic macroinvertebrate assemblage in both of each basin and each season were not significantly different (R = 0.09, p = 0.24 and R = 0.07, p = 0.35, respectively). The efficacy metrics of each basin consisting of the Mekong II, the Chi, and the Mun basins were integrated and calibrated. A total of 255 data sets of water physico-chemical and benthic macroinvertebrates during the dry period (cool and hot seasons) were obtained. The stream classification could be divided into three groups: the reference group (48 stations), the stressed group (42 stations), and the intermediate group (165 stations). Twelve out of 56 metrics have been considered as a core metric for the development of a biological index for quality streams in the Northeast, including Total taxa, EPT taxa, Ephemeroptera taxa, Coleoptera taxa, % EPT, % Chironomidae, % Tolerant individuals, % Intolerant individuals, Beck's index, HBI, Predator taxa, and Clinger taxa. Moreover, this metric set covered the structure and function of organisms including the diversity of species, community structure, tolerance/intolerance measures, functional feeding group, and habit. From the efficacy validation of the biological index, the results of stream assessment corresponded to the classification sites with the physico-chemical characteristics.