Gao S J, Moore P S
School of Public Health, Columbia University New York, New York 10032, USA.
Emerg Infect Dis. 1996 Jul-Sep;2(3):159-67. doi: 10.3201/eid0203.960301.
New molecular biologic techniques, particularly representational difference analysis, consensus sequence-based polymerase chain reaction, and complementary DNA library screening, have led to the identification of several previously unculturable infectious agents. New agents have been found in tissues from patients with Kaposi's sarcoma, non-A, non-B hepatitis, hantavirus pulmonary syndrome, bacillary angiomatosis, and Whipple's disease by using these techniques without direct culture. The new methods rely on identifying subgenomic fragments from the suspected agent. After a unique nucleic acid fragment belonging to an agent is isolated from diseased tissues, the fragment can be sequenced and used as a probe to identify additional infected tissues or obtain extended portions of the agent's genome. For agents that cannot be cultured by standard techniques, these approaches have proved invaluable for identification and characterization studies. Applying these techniques to other human diseases of suspected infectious etiology may rapidly elucidate novel candidate pathogens.
新的分子生物学技术，特别是代表性差异分析、基于共有序列的聚合酶链反应和互补DNA文库筛选，已导致鉴定出几种以前无法培养的感染因子。通过使用这些技术而不进行直接培养，在患有卡波西肉瘤、非甲非乙型肝炎、汉坦病毒肺综合征、杆菌性血管瘤病和惠普尔病患者的组织中发现了新的病原体。这些新方法依赖于从可疑病原体中鉴定亚基因组片段。从患病组织中分离出属于某病原体的独特核酸片段后，该片段可进行测序，并用作探针来鉴定其他受感染组织或获取该病原体基因组的延伸部分。对于无法通过标准技术培养的病原体，这些方法已被证明在鉴定和特征研究中具有极高价值。将这些技术应用于其他疑似感染病因的人类疾病，可能会迅速阐明新的候选病原体。
