Messmer T O, Skelton S K, Moroney J F, Daugharty H, Fields B S
National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, Department of Health and Human Services, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 1997 Aug;35(8):2043-6. doi: 10.1128/jcm.35.8.2043-2046.1997.
We developed a nested, multiplex PCR for simultaneous detection of three species of chlamydiae in human and avian specimens. The PCR was designed to increase sensitivity and to circumvent inhibitors of PCR present in clinical specimens. The target sequence was the 16S rRNA gene. The first-step PCR was genus specific, and the second-step PCR was multiplexed (i.e., had multiple primer sets in the same tube) and could discriminate among Chlamydia pneumoniae, Chlamydia psittaci, and Chlamydia trachomatis on the basis of the molecular weight of the amplicon. The limit of detection of each of the two PCR steps was 5 inclusion-forming units. We used PCR and serologic evidence during outbreaks of psittacosis to infer that C. psittaci had been transmitted from birds purchased in pet stores to humans. We also used this method to test both live and dead birds from pet stores for infection with C. psittaci. Compared with culture, the application of PCR to avian specimens increased the rate of C. psittaci detection.
我们开发了一种巢式多重聚合酶链反应（PCR），用于同时检测人类和禽类标本中的三种衣原体。该PCR旨在提高灵敏度，并规避临床标本中存在的PCR抑制剂。靶序列是16S rRNA基因。第一步PCR是属特异性的，第二步PCR是多重的（即在同一管中有多个引物组），并且可以根据扩增子的分子量区分肺炎衣原体、鹦鹉热衣原体和沙眼衣原体。两个PCR步骤中每个步骤的检测限为5个包涵体形成单位。在鹦鹉热疫情期间，我们利用PCR和血清学证据推断，鹦鹉热衣原体已从宠物店购买的鸟类传播给人类。我们还使用这种方法检测宠物店的活禽和死禽是否感染鹦鹉热衣原体。与培养法相比，将PCR应用于禽类标本提高了鹦鹉热衣原体的检测率。
