Development and evaluation of a rapid recombinase polymerase amplification assay for detection of coxsackievirus A6

Wang, Kaifeng; Wu, Yue; Yin, Dan; Tang, Shixing; Hu, Guifang; He, Yaqing

doi:10.1007/s00705-016-3100-8

Cited by 8 publications

(4 citation statements)

References 21 publications

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“…This study established a highly sensitive and specific diagnostic approach for the detection of SFTSV nucleic acids in clinical serum specimens. The detection limit of SFTSV RT-RPA assay was statistically estimated as 241 copies of target RNA per reaction, which is consistent with other published literature [19][20][21]. While the sensitivity of this RT-RPA assay is lower than conventional RT-qPCR for SFTSV, the detection limit is still far below the viremia level in most clinical SFTS patients [18].…”

Section: Discussionsupporting

confidence: 87%

Development and evaluation of a rapid detection assay for severe fever with thrombocytopenia syndrome virus based on reverse-transcription recombinase polymerase amplification

Zhou

Wang

Zhu

et al. 2020

Molecular and Cellular Probes

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Development and evaluation of a rapid detection assay for severe fever with thrombocytopenia syndrome virus based on reverse-transcription recombinase polymerase amplification, Molecular and Cellular Probes (2020), doi: https://doi.Table of Abbreviations: dT-FAM Thymidine nucleotide carrying FAM fluorophore dT-BHQ1 Thymidine nucleotide carrying BHQ-1 quencher HHV Human herpesvirus LAMP loop-mediated isothermal amplification LOD Limit of detection PIV Parainfluenza virus RPA Recombinase polymerase amplification RT-LAMP Reverse-transcription loop-mediated isothermal amplification RT-PCR Reverse-transcription polymerase chain reaction RT-qPCR Real-time reverse-transcription polymerase chain reaction RT-RAA Reverse-transcription recombinase-aided amplification RT-RPA Real-time reverse-transcription recombinase polymerase amplification SFTS Severe fever with thrombocytopenia syndrome SFTSV Severe fever with thrombocytopenia syndrome virus SSB Single-stranded DNA binding Abstract 1 Rapid detection of severe fever with thrombocytopenia syndrome virus (SFTSV) is 2 crucial for its control and surveillance. In this study, a rapid isothermal real-time 3 reverse-transcription recombinase polymerase amplification (RT-RPA) assay was 4 developed for the detection of SFTSV. The detection limit at 95% probability was 241 5 copies per reaction. A test of 120 serum samples of suspected severe fever with 6 thrombocytopenia syndrome (SFTS) patients revealed that the sensitivity and 7 specificity of the RT-RPA assay was approximately 96.00% (95%CI: 80.46%-99.79%) 8 and 98.95% (95% CI: 94.28%-99.95%), respectively; the kappa value was 0.9495 (P 9 ＜0.001). The Bland-Altman analysis showed that 87.50% of the different data points 10 were located within the 95% limits of agreement, indicating a good correlation 11 between the results from RT-RPA assays and those of RT-qPCR assays. In conclusion, 12 the rapid and efficient RT-RPA assay can be a promising candidate for point-of-care 13 detection method of SFTSV. 14 15 Keywords: Severe fever with thrombocytopenia syndrome virus; Recombinase 16 polymerase amplification; Detection assay Severe fever with thrombocytopenia syndrome virus (SFTSV), belonging to the 19 family Phenuiviridae, the genus Banyangvirus, the species Huaiyangshan 20 banyangvirus, was first identified as the causative pathogen of the outbreak of severe 21 fever with thrombocytopenia syndrome (SFTS) in the central and northeast regions of 22 China in 2009 [1]. During the following decade, this emerging endemic tick-borne 23 disease has been reported in many countries worldwide, including South Korea, Japan, 24 United Arab Emirates, and the United States [2-5]. Delayed admission due to 25 nonspecific clinical manifestations, along with rapid progression to multiple organ 26 dysfunction in severe cases led to a high case fatality rate of 6.40~30% in SFTS 27 patients [6, 7]. 28 Currently, detection methods of SFTSV comprise nucleic acid testing, serological 29 detection, and virus isolation [1, 8, 9]. Due to the long turnaround...

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Section: Discussionsupporting

confidence: 87%

Development and evaluation of a rapid detection assay for severe fever with thrombocytopenia syndrome virus based on reverse-transcription recombinase polymerase amplification

Zhou

Wang

Zhu

et al. 2020

Molecular and Cellular Probes

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“…The analysis suggested comparable performance of RT-LAMP methods to real time RT-PCR, as the pooled sensitivity and specificity were found to be 0.99 (95% CI: 0.97–1.00) vs. 0.97 (95% CI: 0.94–1.00), respectively. More recently, Wang et al (2017) further evaluated this method, finding a limit of detection of 0.01 PFU per reaction. No cross-reaction of the assay was found with poliovirus 1, coxsackievirus A16, rotavirus, norovirus, sapovirus, and astrovirus ( An et al, 2014 ).…”

Section: Small But Deadly: Picornaviridae Familymentioning

confidence: 99%

“…Thus, there is a need to distinguish both for proper treatment. Wang et al (2017) report an RT-RPA assay for coxsackievirus A6 with a detection limit of 400 copies per reaction, with 100% specificity when 234 clinical samples were tested that also included 15 coxsackievirus group A and 5 group B serotypes for which no cross-reactivity was observed. Further, the real-time RT-RPA assay showed no significant difference in sensitivity or specificity with established real-time RT-PCR assays.…”

Section: Small But Deadly: Picornaviridae Familymentioning

confidence: 99%

Recent Developments in Isothermal Amplification Methods for the Detection of Foodborne Viruses

et al. 2022

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Foodborne and enteric viruses continue to impose a significant public health and economic burden globally. As many of these viruses are highly transmissible, the ability to detect them portably, sensitively, and rapidly is critical to reduce their spread. Although still considered a gold standard for detection of these viruses, real time polymerase chain reaction (PCR)-based technologies have limitations such as limited portability, need for extensive sample processing/extraction, and long time to result. In particular, the limitations related to the susceptibility of real time PCR methods to potential inhibitory substances present in food and environmental samples is a continuing challenge, as the need for extensive nucleic acid purification prior to their use compromises the portability and rapidity of such methods. Isothermal amplification methods have been the subject of much investigation for these viruses, as these techniques have been found to be comparable to or better than established PCR-based methods in portability, sensitivity, specificity, rapidity, and simplicity of sample processing. The purpose of this review is to survey and compare reports of these isothermal amplification methods developed for foodborne and enteric viruses, with a special focus on the performance of these methods in the presence of complex matrices.

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“…Recently, recombinase polymerase amplication (RPA) have been emerging as a state-of-the-art, isothermal technology for use in molecular diagnosis of infectious diseases, pathogens and genetically modied organisms. [20][21][22][23][24][25][26][27] Unlike isothermal DNA amplication methods, e.g. Loop-mediated Isothermal AMPli-cation (LAMP) [28][29][30] performed at 60-70 C, PRA reaction proceeds at a temperatures between 37 C and 42 C. 31 RPA process, employing a recombinase enzyme, a single-stranded DNA-binding protein (SSB), a strand-displacing polymerase called Bsu polymerase and two opposing primers, allows exponential amplication of a dened region of DNA molecule.…”

Section: Introductionmentioning

confidence: 99%

Rapid identification of quarantine invasiveSolanum elaeagnifoliumby real-time, isothermal recombinase polymerase amplification assay

Lei

Yan

et al. 2017

RSC Adv.

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Development and evaluation of a rapid recombinase polymerase amplification assay for detection of coxsackievirus A6

Cited by 8 publications

References 21 publications

Development and evaluation of a rapid detection assay for severe fever with thrombocytopenia syndrome virus based on reverse-transcription recombinase polymerase amplification

Development and evaluation of a rapid detection assay for severe fever with thrombocytopenia syndrome virus based on reverse-transcription recombinase polymerase amplification

Recent Developments in Isothermal Amplification Methods for the Detection of Foodborne Viruses

Rapid identification of quarantine invasiveSolanum elaeagnifoliumby real-time, isothermal recombinase polymerase amplification assay

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