Study on Detection of Bacillus licheniformis by Two-site Fluorescence in Situ Hybridization
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Abstract:
In order to investigate the affecting factors of fluorescence in situ hybridization (FISH) on Bacillus, this work used Bacillus licheniformis as its research object. It analyzed the effects of different probes, strain pretreatments, strain permeability treatments, and hybridization conditions on the detection results of FISH, and further detected the specificity of the selected probes. The results showed that two of the designed probes could identify Bacillus licheniformis; specifically, the two probes exhibited a better fluorescence effect than a single probe, having 1.54 times higher pixel intensity. The reaction conditions were the following: ultrasonic dispersing time, 120 s; lysozyme treatment, 40 min; hybridization time, 6 h; formamide concentration 30%. Consequently, the detection result was the best, producing a gray value of 41.16 and a detection rate of 96.35%. The specificity detections determined that the said method could specifically recognize Bacillus licheniformis. A relatively simple two-site fluorescence in situ hybridization approach was established to detect Bacillus licheniformis, with strong specificity without nucleic acid extraction and amplification.
