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基于重组酶聚合酶扩增的鹦鹉热衣原体快速检测方法。

A recombinase polymerase amplification-based assay for rapid detection of Chlamydia psittaci.

机构信息

Guangdong Provincial Animal Virus Vector Vaccine Engineering Technology Research Center, College of Animal Science, South China Agricultural University, Guangzhou 510642, P.R. China.

Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, Guangzhou 510633, P.R. China.

出版信息

Poult Sci. 2021 Feb;100(2):585-591. doi: 10.1016/j.psj.2020.11.031. Epub 2020 Nov 28.

Abstract

Chlamydia psittaci is a zoonotic agent of systemic wasting disease in birds and atypical pneumonia in mammalians including humans, constituting a public health risk. A rapid diagnostic assay would be beneficial in screening C. psittaci in the field. In this study, we developed a probe-based recombinase polymerase amplification (RPA) assay for the rapid detection of C. psittaci. The specific primer pairs and probe targeting the conserved region of the outer membrane protein A gene were designed and applied to the real-time real-time RPA assay. The test can be performed at 39°C for 20 min using a portable device, with sensitivities approaching 100 copies of DNA molecules per reaction, with no cross-reaction with other pathogens. The clinical performance of the RPA assay was evaluated in an outbreak of C. psittaci and has high accuracy levels in field applications. The epidemic C. psittaci strains were classed into 2 genotypes: A and C. Collectively, this study offers a promising approach in screening for C. psittaci both in a laboratory setting and in field settings, and RPA can be used as an effective clinical test to monitor outbreaks in domestic fowl populations.

摘要

鹦鹉热衣原体是一种动物源性全身消耗性疾病的病原体,可导致鸟类发病,同时也是哺乳动物包括人类非典型性肺炎的病原体,对公共卫生构成了威胁。快速诊断检测方法将有利于在现场筛查鹦鹉热衣原体。在本研究中,我们开发了一种基于探针的重组酶聚合酶扩增(RPA)检测方法,用于快速检测鹦鹉热衣原体。设计了针对外膜蛋白 A 基因保守区的特异性引物对和探针,并应用于实时实时 RPA 检测。该检测可在 39°C 下进行 20 分钟,使用便携式设备,反应的灵敏度接近 100 拷贝 DNA 分子,与其他病原体无交叉反应。RPA 检测方法的临床性能在鹦鹉热衣原体爆发中进行了评估,在现场应用中具有较高的准确度。流行的鹦鹉热衣原体菌株分为 2 个基因型:A 和 C。总之,本研究为在实验室和现场筛查鹦鹉热衣原体提供了一种有前途的方法,RPA 可作为监测家禽群体中爆发的有效临床检测方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e33/7858173/1cacf940f83b/gr1.jpg

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