Improving the molecular diagnosis of Chlamydia psittaci and Chlamydia abortus infection with a species-specific duplex real-time PCR
Improving the molecular diagnosis of Chlamydia psittaci and Chlamydia abortus infection with a species-specific duplex real-time PCR
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DOI：
10.1099/jmm.0.000139
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发表时间：
2015-10-01
影响因子：
3
通讯作者：
Greub, Gilbert
中科院分区：
文献类型：
--
作者：
Opota, Onya;Jaton, Katia;Greub, Gilbert
鹦鹉热衣原体和流产衣原体是密切相关的胞内细菌，表现出不同的组织趋向性，可能导致严重但不同的人类感染。鹦鹉热会引起鹦鹉热，这是一种由鸟类传播的呼吸道人畜共患传染病。流产梭菌是一种小型反刍动物的引产因子，也可定植于人的胎盘，导致胎儿死亡和流产。鹦鹉热杆菌和流产弧菌引起的感染被低估，主要是因为它们严格的细胞内生长导致诊断困难。我们建立了一种检测和区分临床标本中这两种细菌的双重实时荧光聚合酶链式反应方法。第一次聚合酶链式反应(PCR1)针对16S-23S rRNA操纵子序列，可同时检测鹦鹉热链球菌和流产弧菌。第二次聚合酶链式反应(PCR2)针对鹦鹉热虫特有的编码DNA序列CPSIT_0607。两次聚合酶链式反应均可100%检出10个DNA拷贝(1000个拷贝ml(-1))。利用120份样本，包括细菌参考菌株、临床标本和感染细胞培养材料，我们监测了检测鹦鹉热弧菌和流产弧菌PCR1的100%敏感性和100%特异性。当PCR1阳性时，PCR2可区分鹦鹉热链球菌和流产弧菌，其阳性预测值为100%，阴性预测值为88%。总之，这种新的双重聚合酶链式反应是一种低成本、节省时间、具有高通量潜力的方法，有望提高大规模筛查和暴发期间鹦鹉热病和妊娠并发症的常规诊断。
Chlamydia psittaci and Chlamydia abortus are closely related intracellular bacteria exhibiting different tissue tropism that may cause severe but distinct infection in humans. C. psittaci causes psittacosis, a respiratory zoonotic infection transmitted by birds. C. abortus is an abortigenic agent in small ruminants, which can also colonize the human placenta and lead to foetal death and miscarriage. Infections caused by C. psittaci and C. abortus are underestimated mainly due to diagnosis difficulties resulting from their strict intracellular growth. We developed a duplex real-time PCR to detect and distinguish these two bacteria in clinical samples. The first PCR (PCR1) targeted a sequence of the 16S-23S rRNA operon allowing the detection of both C. psittaci and C. abortus. The second PCR (PCR2) targeted the coding DNA sequence CPSIT_0607 unique to C. psittaci. The two PCRs showed 100 % detection for >= 10 DNA copies per reaction (1000 copies ml(-1)). Using a set of 120 samples, including bacterial reference strains, clinical specimens and infected cell culture material, we monitored 100 % sensitivity and 100 % specificity for the detection of C. psittaci and C. abortus for PCR1. When PCR1 was positive, PCR2 could discriminate C. psittaci from C. abortus with a positive predictive value of 100 % and a negative predictive value of 88 %. In conclusion, this new duplex PCR represents a low-cost and time-saving method with high-throughput potential, expected to improve the routine diagnosis of psittacosis and pregnancy complication in large-scale screening programs and also during outbreaks.