Use of a rapid reverse-transcription recombinase aided amplification assay for respiratory syncytial virus detection

Chen, Chen; Li, Xinna; Li, Guixia; Zhao, Li; Duan, Su-xia; Yan, Teng-fei; Feng, Zhishan; Ma, Xuejun

doi:10.1016/j.diagmicrobio.2017.10.005

Cited by 52 publications

(39 citation statements)

References 33 publications

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“…Moreover, the rapid reverse‐transcription recombinase‐aided amplification (RT‐RAA) assay was developed as a molecular‐based diagnostic method to detect subgroup RSV A and B genomes in clinical specimens. This method mainly utilizes an enzyme mixture, including single‐strand DNA binding protein (SSB), recombinase UvsX, and DNA polymerase, to detect RNA amplicons of RSV 26 . It is performed at 39°C in less than 30 minutes with high specificity.…”

Section: Human Respiratory Syncytial Virus and Its Diagnostic Approachesmentioning

confidence: 99%

Recent advances in the detection of respiratory virus infection in humans

Zhang

Wang

Deng

et al. 2020

Journal of Medical Virology

445

363

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Respiratory tract viral infection caused by viruses or bacteria isone of the most common diseases in human worldwide, while those caused by emerging viruses, such as the novel coronavirus, 2019-nCoV that caused the pneumonia outbreak in Wuhan, China most recently, have posed great threats to global public health. Identification of the causative viral pathogens of respiratory tract viral infections is important to select an appropriate treatment, save people's lives, stop the epidemics, and avoid unnecessary use of antibiotics. Conventional diagnostic tests, such as the assays for rapid detection of antiviral antibodies or viral antigens, are widely used in many clinical laboratories. With the development of modern technologies, new diagnostic strategies, including multiplex nucleic acid amplification and microarray-based assays, are emerging. This review summarizes currently available and novel emerging diagnostic methods for the detection of common respiratory viruses, such as influenza virus, human respiratory syncytial virus, coronavirus, human adenovirus, and human rhinovirus. Multiplex assays for simultaneous detection of multiple respiratory viruses are also described. It is anticipated that such data will assist researchers and clinicians to develop appropriate diagnostic strategies for timely and effective detection of respiratory virus infections. K E Y W O R D S adenovirus, coronavirus, diagnostic methods, influenza virus, respiratory syncytial virus, respiratory viral infection, rhinovirus

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Section: Human Respiratory Syncytial Virus and Its Diagnostic Approachesmentioning

confidence: 99%

Recent advances in the detection of respiratory virus infection in humans

Zhang

Wang

Deng

et al. 2020

Journal of Medical Virology

445

363

View full text Add to dashboard Cite

show abstract

“…Thus, a reliable detection method is critical for clinical diagnosis. In our previous studies, the development of RAA assays for the detection of several pathogens, such as the RSV-A and -B [27], coxsackievirus A10 and coxsackievirus A6 [28], and single-nucleotide polymorphisms (SNPs) [29] only focused on detection of pathogens but did not successfully apply an internal control to monitor the reaction system. Since the RAA reaction is an enzymatic reaction controlled by a variety of enzymes, it is more susceptible to the influence of inhibitors in the specimen.…”

mentioning

confidence: 99%

Development of a duplex reverse transcription recombinase-aided amplification assay for respiratory syncytial virus incorporating an internal control

Zhang

et al. 2019

Arch Virol

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Human respiratory syncytial virus (RSV) is a common viral pathogen that causes lower respiratory tract infections in infants and children globally. In this study, we developed a duplex reverse transcription recombinase-aided amplification (duplex-rtRAA) assay containing an internal control in a single closed tube for the detection of human RSV. The internal control in the amplification effectively eliminated false-negative results and ensured the accuracy of the duplex-rtRAA system. We first developed and evaluated a universal singleplex-rtRAA assay for RSV. The sensitivity of this assay for RSV was determined as 4.4 copies per reaction, and the specificity was 100%. Next, a duplex-rtRAA assay with an internal control was established. The sensitivity of the duplex-rtRAA assay approached 5.0 copies per reaction, and no cross-reaction with other common respiratory viruses was observed. The two detection methods (singleplex-rtRAA and duplex-rtRAA) developed in this study were used to test 278 clinical specimens, and the results showed absolute consistency with RSV RT-qPCR analysis, demonstrating 100% diagnostic sensitivity and specificity. These data indicate that the duplex-rtRAA has great potential for the rapid detection of RSV with a high sensitivity.Acute respiratory tract infections (ARTIs) are one of the main causes of outpatient visits, hospitalization, and death, especially in children, the elderly and immunocompromised individuals [1-3]. The major pathogens causing ARTIs in infants and young children are viruses, the most frequently reported of which include respiratory syncytial virus (RSV),

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“…Recombinase‐aided amplification can also use reverse transcriptase and a fluorescent probe system to detect RNA amplicons in real time (Zhang et al, ). Until now, RAA has only been used for detecting human pathogens, including Salmonella (Zhang et al, ), respiratory syncytial virus (Chen et al, ; Qi et al, ), coxsackievirus (Yan et al, ), hepatitis B virus (Shen et al, ) and Schistosoma japonicum ‐specific gene fragments,(Song et al, ) and there have been no previous reports of the use of RAA for detecting animal pathogens. Thus, the current study provides the first report of an H7 RT‐RAA assay for the detection of an animal pathogen.…”

Section: Discussionmentioning

confidence: 99%

“…Recombinase‐aided amplification is a rapid, simple method that can be completed in 20 min at approximately 39°C. Recombinase‐aided amplification has already been successfully used to detect bacterial (Zhang et al, ) and viral pathogens (Chen et al, ; Yan et al, ).…”

Section: Introductionmentioning

confidence: 99%

Reverse‐transcription recombinase‐aided amplification assay for H7 subtype avian influenza virus

Wang

Huang

Liu

et al. 2019

Transbound Emerg Dis

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H7 subtype avian influenza virus infection is an emerging zoonosis in some Asian countries and an important avian disease worldwide. A rapid and simple test is needed to confirm infection in suspected cases during disease outbreaks. In this study, we developed a reverse‐transcription recombinase‐aided amplification assay for the detection of H7 subtype avian influenza virus. Assays were performed at a single temperature (39°C), and the results were obtained within 20 min. The assay showed no cross‐detection with Newcastle disease virus or infectious bronchitis virus, which are the other main respiratory viruses affecting birds. The analytical sensitivity was 102 RNA copies per reaction at a 95% probability level according to probit regression analysis, with 100% specificity. Compared with published reverse‐transcription quantitative real‐time polymerase chain reaction assays, the κ value of the reverse‐transcription recombinase‐aided amplification assay in 342 avian clinical samples was 0.988 (p < .001). The sensitivity for avian clinical sample detection was 100% (95%CI, 90.40%–100%), and the specificity was 99.96% (95%CI, 97.83%–99.98%). These results indicated that our reverse‐transcription recombinase‐aided amplification assay may be a valuable tool for detecting avian influenza H7 subtype virus.

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Use of a rapid reverse-transcription recombinase aided amplification assay for respiratory syncytial virus detection

Cited by 52 publications

References 33 publications

Recent advances in the detection of respiratory virus infection in humans

Recent advances in the detection of respiratory virus infection in humans

Development of a duplex reverse transcription recombinase-aided amplification assay for respiratory syncytial virus incorporating an internal control

Reverse‐transcription recombinase‐aided amplification assay for H7 subtype avian influenza virus

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