Piémont Y, Jaulhac B
Institut de Bactérologie de la Faculté de Médecine, Université Louis-Pasteur, Strasbourg.
Ann Dermatol Venereol. 1995;122(4):206-12.
Progress in molecular biology has led to the development of new tools for bacteriological diagnosis. Sporadic genes coding for virulence factors can be detected with highly specific genetic probes applied to cultured bacteria. Such genetic probes can also be used to specifically identified cultured bacteria whose general taxonomic classification is known. Another advantage of molecular genetics is the possibility that the cell culture step may not be needed, bacteria being identified directly in the sample specimen. Such techniques are particularly interesting to identify bacteria which are difficult to culture (for example: Borrelia burgdorferi, Chlamydia trachomatis) or which grow slowly (mycobacteria). The bacterial DNA must be isolated and amplified with an enzyme reaction. This is a critical step in the method: several positive and negative controls are required. When performed under optimal conditions, amplification techniques are excellent methods which can offer results similar to culture methods in culturable bacteria. Finally, molecular biology can be used to identify previously cultured bacteria for which there is no taxonomic orientation. Here the ribosome 165 DNA must be amplified and sequenced. The sequence is then compared with a data bank allowing classification. One could image future techniques applied to certain pathology samples for the detection and identification of bacteria without need for a culture step. However, direct microscope examination and bacterial culture remain the basic methods for bacteriologic diagnosis, the advantages and disadvantages of molecular biology leading to its use a complementary method for improving the quality of the diagnosis.
分子生物学的进展带来了细菌学诊断新工具的发展。编码毒力因子的散在基因可通过应用于培养细菌的高度特异性基因探针进行检测。此类基因探针还可用于特异性鉴定已知一般分类学分类的培养细菌。分子遗传学的另一个优势是有可能无需细胞培养步骤,直接在样本标本中鉴定细菌。此类技术对于鉴定难以培养的细菌(例如:伯氏疏螺旋体、沙眼衣原体)或生长缓慢的细菌(分枝杆菌)特别有意义。必须通过酶反应分离并扩增细菌DNA。这是该方法中的关键步骤:需要几个阳性和阴性对照。在最佳条件下进行时,扩增技术是优秀的方法,对于可培养细菌能提供与培养方法类似的结果。最后,分子生物学可用于鉴定尚无分类学定位的先前培养细菌。在此必须扩增核糖体165 DNA并进行测序。然后将序列与允许分类的数据库进行比较。可以设想未来应用于某些病理样本的技术,无需培养步骤即可检测和鉴定细菌。然而,直接显微镜检查和细菌培养仍然是细菌学诊断的基本方法,分子生物学的优缺点使其成为提高诊断质量的补充方法。