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本研究利用脆弱类杆菌、产黑素类杆菌和核梭杆菌经碱裂解法抽提全染色体 DNA,并以缺口移位法标记上~(32)P 以制备探针,对37株感染根管内厌氧菌进行了斑点杂交,根据X线片上显影结果判断杂交率以鉴定细菌。结果与常规鉴定基本吻合,显示出很高的特异性和敏感性,并具有快速简便的优点。交叉反应现象只表现出弱阳性杂交信号,可以与特异性杂交相区别,对此可通过改进杂交和冲洗条件或使用寡核苷酸探针而加以完善。
In this study, whole-body DNA was extracted by alkaline lysis using Bacteroides fragilis, Melanocytes and Fusobacterium nucleatum, and ~ (32) P was labeled by gap-shift method to prepare probes. Within the anaerobic bacteria were spotted hybridization, X-ray on the basis of the results of development to determine the rate of hybridization to identify bacteria. The result is basically consistent with the routine identification, showing high specificity and sensitivity, and has the advantages of quickness and simplicity. Cross-reaction phenomena show only weakly positive hybridization signals that can be distinguished from specific hybridization, which can be improved by improving hybridization and washout conditions or by using oligonucleotide probes.