Study On Methods Of Detecting Bacillus Atrophaeus

Bacillus atrophaeus, a variant of Bacillus subtilis, which forms a soluble black pigment on media containing utilizable carbohydrates. At present, studies at home and abroad dedicated to its application research, but little research on the bacteria detection method. Therefore, there was no mature or easy method for reference. From the contraction of bacillus biological characteristics, this article do a system research on this bacteria, to explore a rapid, accurate and reliable biological detect method,which have a great significance for its wider and more systematic application in the future.According to the detect method of bacillus, this study used ATCC9372which is widely used in disinfection and sterilization area as representative strains of Bacillus atrophaeus for research, puts forward the declining physical chemical method and PCR technique method to detect Bacillus atrophaeus.First of all, according to the physiological experimental study, summarized the physiological characteristics of Bacillus atrophaeus, proposed the traditional detect method of Bacillus atrophaeus. select bacillus on microscopy first, then use the API50CHB test strip to test the ability of metabolism, add tests including anaerobic growth, nitrate utilization, V.P test, starch decomposition, D-xylose utilization, L-Arab sugar utilization, utilization of D-mannitol,10%NaCl growth as confirmed project.Second, in terms of molecular detection, the PCR methods were used to detect Bacillus atrophaeus. By screening species-specific primers (recA and ITS primer) and optimization annealing temperature, determined the appropriate amplification conditions.Results showed that the Bacillus atrophaeus can produced131stripe to recA primer and75bp stripe to ITS primer, while other9kinds of bacillus species had none specific amplification.Good specificity and sensitivity were demonstrated for these methods. At last, validation test of Bacillus atrophaeus were taken on environmental samples by physiological detection method and PCR technology method. Both methods had good detection effect,physiological method has low technical requirements, wide applicability, but time-consuming, troublesome, and the result is not stable, easily effected by bacteria content and environment changes; PCR identification method is accurate, fast, strong specificity, but instruments and technical requirements are relatively high.To sum up, this paper successfully established the detection method of Bacillus atrophaeus, including physiological identification method and PCR detection method, provides convenience to Bacillus atrophaeus’ further research and development.