Eboigbodin Kevin E, Moilanen Kirsi, Elf Sonja, Hoser Mark
Research and Development, Orion Diagnostica Oy, P. O. BOX 83, FI-02101, Espoo, Finland.
Molecular Biology, GeneForm Technologies, Broadstairs, UK.
BMC Infect Dis. 2017 Feb 10;17(1):134. doi: 10.1186/s12879-017-2227-x.
Respiratory syncytial virus (RSV) is one of the most common causes of respiratory tract infections among young children and the elderly. Timely and accurate diagnosis of respiratory tract infections improves patient care and minimizes unnecessary prescriptions of antibiotics. We sought to develop a rapid nucleic acid tests for the detection of RSV within minutes, while retaining the high sensitivity achieved with RT-PCR.
We developed and evaluated a reverse transcription isothermal nucleic acid amplification method, reverse transcription strand invasion based amplification (RT-SIBA), for the rapid detection of RSV.
The developed RT-SIBA assay showed good sensitivity by detecting as few as 10 copies of RSV RNA within 20 min compared with reverse transcription polymerase chain reaction, which took approximately 2 h. The performance of the RT-SIBA RSV assay was further investigated using nasopharyngeal swab specimens. The RT-SIBA assay had a sensitivity of 100% (25/25) and a specificity of 100% (15/15).
RT-SIBA did not require highly purified RNA for the rapid detection of RSV and was therefore compatible with rapid specimen processing methods. This reduces the complexity of specimen preparation and further shortens the total amount of time needed to detect RSV in clinical specimens. The developed RT-SIBA assay for RSV could be a useful tool for prompt management of this infection.
呼吸道合胞病毒(RSV)是幼儿和老年人呼吸道感染最常见的病因之一。及时准确地诊断呼吸道感染可改善患者护理,并尽量减少不必要的抗生素处方。我们试图开发一种能在几分钟内检测RSV的快速核酸检测方法,同时保持逆转录聚合酶链反应(RT-PCR)所具有的高灵敏度。
我们开发并评估了一种用于快速检测RSV的逆转录等温核酸扩增方法,即基于链侵入的逆转录扩增法(RT-SIBA)。
与耗时约2小时的RT-PCR相比,所开发的RT-SIBA检测方法在20分钟内就能检测到低至10拷贝的RSV RNA,显示出良好的灵敏度。使用鼻咽拭子标本进一步研究了RT-SIBA RSV检测方法的性能。RT-SIBA检测方法的灵敏度为100%(25/25),特异性为100%(15/15)。
RT-SIBA在快速检测RSV时不需要高度纯化的RNA,因此与快速标本处理方法兼容。这降低了标本制备的复杂性,并进一步缩短了临床标本中检测RSV所需的总时间。所开发的用于检测RSV的RT-SIBA检测方法可能是及时管理这种感染的有用工具。