Establishment of an Analysis Method for Intestinal Flora Dysregulation in Mice Based on an ERIC-PCR Fingerprinting Technique

Yong-Zhi LUN, Jie SUN, Zhi-Peng RUAN, Ling-Hong PAN

Abstract


A conventional flora analysis method, as well as an ERIC-PCR technique, were employed to detect the intestinal microflora of antibiotic associated diarrhea in a mice model and normal mice, respectively. Then, combined with the results of the bacterial cultures and DNA fingerprint detection, the changes in the dominant intestinal flora quantities and species of the mice were analyzed. Also, a detection method which utilized ERIC-PCR technology for the analysis of the imbalances in the intestinal flora of the mice was established. First, a conventional analysis method was applied for the identification of the bacteria in normal mice, and mice with antibiotic associated diarrhea flora. Then, the genomic DNA was extracted. Finally, an enterobacterial repetitive intergenic sequence (ERIC) method was employed to obtain the intestinal microflora fingerprint of the normal mice, and the model mice using ERIC as the template. The accuracy of the ERIC-PCR technology was verified by comparing with the results of the analysis of the conventional bacteria group. The conventional flora analysis results showed that there were significant changes in the number of the four types of dominant bacteria, which confirmed that the model had been successfully constructed. The feces DNA genome maps of the mice in the two groups were successfully obtained using the ERIC-PCR. Moreover, a specific fingerprint with a certain contrast was found to be exhibited in the two groups. From the mouse feces genomes obtained using the ERIC-PCR, it could be observed that there were specific bands of distribution, number, and brightness of the genome. These results indicated that there were differences in the distributions of the intestinal flora. When combined with the conventional flora analysis method comparison, it could be seen that the ERIC-PCR technique was an efficient detection method for the analysis of the intestinal flora dysregulation of the mice.


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