用于检测牙龈卟啉单胞菌和福赛斯坦纳菌的随机克隆DNA探针与全基因组DNA探针的比较
Comparison of randomly cloned and whole genomic DNA probes for the detection of Porphyromonas gingivalis and Bacteroides forsythus.
作者信息
Wong M, DiRienzo J M, Lai C H, Listgarten M A
机构信息
Department of Periodontics, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104-6002, USA.
出版信息
J Periodontal Res. 1996 Jan;31(1):27-35. doi: 10.1111/j.1600-0765.1996.tb00460.x.
Whole genomic and randomly-cloned DNA probes for two fastidious periodontal pathogens, Porphyromonas gingivalis and Bacteroides forsythus were labeled with digoxigenin and detected by a colorimetric method. The specificity and sensitivity of the whole genomic and cloned probes were compared. The cloned probes were highly specific compared to the whole genomic probes. A significant degree of cross-reactivity with Bacteroides species, Capnocytophaga sp. and Prevotella sp. was observed with the whole genomic probes. The cloned probes were less sensitive than the whole genomic probes and required at least 10(6) target cells or a minimum of 10 ng of target DNA to be detected during hybridization. Although a ten-fold increase in sensitivity was obtained with the whole genomic probes, cross-hybridization to closely related species limits their reliability in identifying target bacteria in subgingival plaque samples.
用洋地黄毒苷标记了针对两种苛求性牙周病原体——牙龈卟啉单胞菌和福赛坦氏菌的全基因组和随机克隆的DNA探针，并采用比色法进行检测。比较了全基因组探针和克隆探针的特异性和敏感性。与全基因组探针相比，克隆探针具有高度特异性。全基因组探针与拟杆菌属、二氧化碳嗜纤维菌属和普雷沃菌属存在显著程度的交叉反应。克隆探针的敏感性低于全基因组探针，在杂交过程中需要至少10⁶个靶细胞或至少10 ng靶DNA才能被检测到。尽管全基因组探针的敏感性提高了10倍，但与密切相关物种的交叉杂交限制了它们在鉴定龈下菌斑样本中靶细菌时的可靠性。
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