Rapid and sensitive real-time assay for the detection of respiratory syncytial virus using RT-SIBA®

Abstract

Background: 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.

Methods: 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.

Results: 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).

Conclusion: 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.

Fig. 1

Sensitivity of RT-SIBA for the detection of respiratory syncytial virus (RSV). a RSV-A RNA and b RSV-B RNA. c Melting curve profiles of RSV (1000 copies of RSV-A RNA) and MS2 RNA in the same reaction tube. MS2 was used as the internal control (IC, 2000 copies per reaction). No template control (NTC)