Rapid and visual detection of human enterovirus coxsackievirus A16 by reverse transcription loop-mediated isothermal amplification combined with lateral flow device

Guo, Yan; Liu, Jun; Zhao, Keyan; Ge, Yiyue; Yang, Yongqin; Zhang, Xiaoyu; Shi, Zhiyang; Cui, Lunbiao

doi:10.1111/lam.12499

Cited by 13 publications

(5 citation statements)

References 31 publications

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“…Biotinstreptavidin interaction as one of the strongest noncovalent interactions has some unique characteristics that make it a very good 'bridge system' in many applications including nucleic acid hybridization assays (Chivers et al, 2011;Mukama et al, 2020). The same principle of biotin and streptavidin molecules interaction concept was also applied in other LAMP-LFD assays such as for the detection of classical swine fever virus (Chowdry et al, 2014), human enterovirus coxsackievirus (Yan et al, 2015) and Staphylococcus aureus (Nawattanapaiboon et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Loop-mediated isothermal amplification of the toxR gene coupled with lateral flow dipstick (LAMP-LFD) for the novel, rapid and specific visual detection of Vibrio harveyi

Rahman, A. M. A.,

Ransangan, J.,

Subbiah, V. K.

2023

MJM

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Aims: Vibrio harveyi is a serious pathogen for marine organisms particularly in hatcheries and grow-out ponds that attack their immune system. The rapid detection of V. harveyi is urgently needed to prevent bacterial spread. Here we described a rapid and specific visual detection method based on the Loop-Mediated Isothermal Amplification in combination with Lateral Flow Dipstick (LAMP-LFD). Methodology and results: A set of six novel primers were designed to target the toxR gene. These include the biotinlabelled inner primer that complements specifically to the target sequences. The resulting biotinylated LAMP amplicons were hybridised to the FAM-labelled probe resulting in lateral flow detection on the dipstick. The addition of loop primers improved the reaction time of LAMP by more than half and rapid detection was observed within 10-15 min. In comparison, the sensitivity of PCR-UV analysis was only at 10 4 copies while LAMP-LFD was able to detect lower amounts at 10 3 copies. The LFD provided higher specificity and selectivity since hybridization with specific probes to the LAMP amplicons was employed. In addition, detection of V. harveyi infected grouper was successful using the LAMP-LFD method described here. Conclusion, significance and impact of study: LAMP-LFD is specific to V. harveyi. Our method provides a useful tool to rapidly detect and monitor the outbreaks of the pathogen.

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Section: Discussionmentioning

confidence: 99%

Loop-mediated isothermal amplification of the toxR gene coupled with lateral flow dipstick (LAMP-LFD) for the novel, rapid and specific visual detection of Vibrio harveyi

Rahman, A. M. A.,

Ransangan, J.,

Subbiah, V. K.

2023

MJM

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“…One of such modification is Immunocapture (IC)-LAMP, where canine Parvovirus was first captured by surface antigen specific antibody followed by isothermal amplification of its genome or IC-LAMP with ELISA where biotin-labeled probes were designed for hybridization with LAMP amplicons on streptavidin-coated wells for visual confirmation of canine parvovirus (Figure 3; Sun et al, 2014, 2017). Another modification is LAMP- lateral flow dipstick (LAMP-LFD) which utilizes FITC labeled probes for rapid and visual detection of canine Parvovirus and human Enterovirus, Coxsackie virus (Sun et al, 2014; Yan et al, 2015). The sensitivity and simplicity of these methods were much superior as compared to conventional PCR.…”

Section: Enteric Virus Detection Methodsmentioning

confidence: 99%

Advances in Diagnostic Approaches for Viral Etiologies of Diarrhea: From the Lab to the Field

et al. 2019

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The applications of correct diagnostic approaches play a decisive role in timely containment of infectious diseases spread and mitigation of public health risks. Nevertheless, there is a need to update the diagnostics regularly to capture the new, emergent, and highly divergent viruses. Acute gastroenteritis of viral origin has been identified as a significant cause of mortality across the globe, with the more serious consequences seen at the extremes of age groups (young and elderly) and immune-compromised individuals. Therefore, significant advancements and efforts have been put in the development of enteric virus diagnostics to meet the WHO ASSURED criteria as a benchmark over the years. The Enzyme-Linked Immunosorbent (ELISA) and Polymerase Chain Reaction (PCR) are the basic assays that provided the platform for development of several efficient diagnostics such as real-time RT-PCR, loop-mediated isothermal amplification (LAMP), polymerase spiral reaction (PSR), biosensors, microarrays and next generation sequencing. Herein, we describe and discuss the applications of these advanced technologies in context to enteric virus detection by delineating their features, advantages and limitations.

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“…EV71 viruses were used with zero cross reactivity between them, suggesting strong specificity; however, other subtypes of coxsackieviruses were not tested. A similar study combined RT-LAMP technology with a lateral flow device, which had a sensitivity of 0.55 TCID50 per reaction and 100% specificity in detecting coxsackievirus A16 ( Yan et al, 2015 ).…”

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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Rapid and visual detection of human enterovirus coxsackievirus A16 by reverse transcription loop-mediated isothermal amplification combined with lateral flow device

Cited by 13 publications

References 31 publications

Loop-mediated isothermal amplification of the toxR gene coupled with lateral flow dipstick (LAMP-LFD) for the novel, rapid and specific visual detection of Vibrio harveyi

Loop-mediated isothermal amplification of the toxR gene coupled with lateral flow dipstick (LAMP-LFD) for the novel, rapid and specific visual detection of Vibrio harveyi

Advances in Diagnostic Approaches for Viral Etiologies of Diarrhea: From the Lab to the Field

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

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