Evaluation and application of reverse transcription loop‐mediated isothermal amplification for detection of noroviruses

Yoda, Tomoko; Suzuki, Yasuhiko; Yamazaki, Kenji; Sakon, Naomi; Kanki, Masashi; Aoyama, Ikuko; Tsukamoto, Teizo

doi:10.1002/jmv.20802

Cited by 64 publications

(41 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Compared to conventional RT-PCR (184), the RT-LAMP assay had 100% clinical sensitivity. Based on this work, a commercial assay, Loopamp Norovirus GI and GII, was developed (Eiken Chemical, Tokyo, Japan) (232). One advantage of this approach is that detection is performed via inexpensive, real-time turbidimetry or simple, endpoint, visual examination.…”

Section: Molecular Diagnostic Testsmentioning

confidence: 99%

Norovirus

2015

View full text Add to dashboard Cite

SUMMARY Norovirus, an RNA virus of the family Caliciviridae , is a human enteric pathogen that causes substantial morbidity across both health care and community settings. Several factors enhance the transmissibility of norovirus, including the small inoculum required to produce infection (<100 viral particles), prolonged viral shedding, and its ability to survive in the environment. In this review, we describe the basic virology and immunology of noroviruses, the clinical disease resulting from infection and its diagnosis and management, as well as host and pathogen factors that complicate vaccine development. Additionally, we discuss overall epidemiology, infection control strategies, and global reporting efforts aimed at controlling this worldwide cause of acute gastroenteritis. Prompt implementation of infection control measures remains the mainstay of norovirus outbreak management.

show abstract

Section: Molecular Diagnostic Testsmentioning

confidence: 99%

Norovirus

2015

View full text Add to dashboard Cite

show abstract

“…The loop-mediated isothermal amplification (LAMP) method has been shown to be highly accurate for the detection of DNA (4,20) and RNA (14,32,36) viruses, differentiation of viral serotypes and subtypes (17,21), and rapid diagnosis of bacterial infections (5). LAMP amplifies target nucleic acid under isother-mal conditions, usually between 60°C and 65°C (19).…”

mentioning

confidence: 99%

Development and Evaluation of a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens

et al. 2009

View full text Add to dashboard Cite

This paper reports on the development and validation of a real-time reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) targeting the genomic large RNA segment of Rift Valley fever virus (RVFV). The set of six designed RT-LAMP primers identified strains of RVFV isolated in geographically distinct areas over a period of 50 years; there was no cross-reactivity with other genetically related and unrelated arboviruses. When testing serial sera and plasma from sheep experimentally infected with wild-type RVFV, there was 100% agreement between results of the RT-LAMP, a TaqMan-based real-time PCR, and virus isolation. Similarly, the assay had very high levels of diagnostic sensitivity and specificity when testing various clinical specimens from humans and animals naturally infected with the virus during recent outbreaks of the disease in Africa. The detection of specific viral genome targets in positive clinical specimens was achieved in less than 30 min. As a highly accurate, rapid, and very simple nucleic acid detection format, the RT-LAMP has the potential to be used in less-well-equipped laboratories in Africa and as a portable device during RVF outbreaks in remote areas, and it can be a valuable tool for the differential diagnosis of viral hemorrhagic fevers.

show abstract

Section: Strategies To Miniaturize Isothermal Amplification Reactionsmentioning

confidence: 99%

“…Calcein has been used for the real-time detection of DNA formation during LAMP [46]. Further methods apply intercalating DNA dyes such as SYBR Green I [47], FDR [48], or oligonucleotide probes labeled with different fluorescent markers as well as low molecular weight cationic polymers such as polyethylenimine [49].To perform the reaction, a set of two specially designed inner and outer primer pairs and a DNA polymerase with strand displacement activity are required for the DNA synthesis. The initial reaction steps are illustrated in Figure 1.…”

mentioning

confidence: 99%

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Tröger¹,

Niemann²,

Gärtig³

et al. 2015

J Nanomed Nanotechnol

View full text Add to dashboard Cite

Nucleic acid amplification technologies (NAATs) offer the most sensitive tests in the clinical laboratory. These techniques are used as a powerful tool for screening and diagnosis of infectious diseases. Isothermal methods, as an alternative to polymerase chain reaction (PCR), require no thermocycling machine and can mostly be performed with reduced time, high throughput, and accurate and reliable results.However, current molecular diagnostic approaches generally need manual analysis by qualified and experienced personal which is a highly complex, time-consuming and labor-intensive task. Thus, the demand for simpler, miniaturized systems and assays for pathogen detection is steadily increasing. Microfluidic platforms and lab-on-a-chip devices have many advantages such as small sample volume, portability and rapid detection time and enable point-of-care diagnosis.In this article, we review several isothermal amplification methods and their implementation in microsystems in relation to quantification of nucleic acids. miniaturized systems can be diverse. The majority of publications are focussed on clinical diagnostics including foodborne organisms for environmental monitoring [10][11][12]. In case of an infectious disease, cultivation and phenotypic characterization are still the standard methods of microbiologists which need days up to weeks to obtain a diagnostic result. Hence, scientists started to make use of nucleic acid amplification methods to accelerate the analysis. The most widespread and well known technique for this purpose is the polymerase chain reaction (PCR) [10,13], which exploits the activity of the DNA polymerase. The method is based on a thermal cycling process consisting of repeated cycles of heating and cooling steps allowing for DNA melting, sequence specific primer binding and enzymatic amplification of the target nucleic acid. The quantitative assessment of target copies is possible by using a real-time PCR approach, where a concentration dependent signal (e.g. fluorescence) increases over time [14]. The latter enables the user to easily assess the pathogen load of patient samples [15][16][17][18]. Since the first example of a PCR on a miniaturized system in 1994 [19,20], numerous devices were presented, which integrate not just the amplification, but also sample preparation and detection steps [9,[21][22][23][24][25][26][27][28]. Just a few of those microsystems have reached the market so far, but some have shown a fascinating potential to replace the classical laboratory-oriented state-of-the art [29][30][31][32][33]. setups has been shown to obtain a similar result [34]. A smaller heat capacity allows for rapid changes in temperature beneficial for the PCR time as well as a higher parallelism of multiple genetic samples [34].However, a major drawback of the integration of PCR to microsystems is the necessity of sophisticated instrumentation. The reaction requires a thermal cycling instrumentation, space and considerable expertise [35]. Therefore, isothermal reactions for the ta...

show abstract

Evaluation and application of reverse transcription loop‐mediated isothermal amplification for detection of noroviruses

Cited by 64 publications

References 21 publications

Norovirus

Norovirus

Development and Evaluation of a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Contact Info

Product

Resources

About