Colorimetric detection of norovirus by helicase-dependent amplification method based on specific primers integrated with HRPzyme

Lee, Jeong Eun; Kim, Sol-A; Park, Hyunjin; Mun, Hyoyoung; Ha, Kwang-Soo; Shim, Won‐Bo

doi:10.1007/s00216-022-04247-5

Cited by 8 publications

(9 citation statements)

References 28 publications

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“…3 B illustrates the procedural overview of the HDA technique. The process starts with thermal denaturation of the target dsDNA or RNA, typically at a high temperature around 90 °C ( Lee et al, 2022a , Moon et al, 2022 ). The reaction temperature is then adjusted to allow primer annealing, typically within 37 °C to 65 °C.…”

Section: Recent Advances In Naa-based Methodsmentioning

confidence: 99%

“…The reaction temperature is then adjusted to allow primer annealing, typically within 37 °C to 65 °C. Successful primer annealing precedes the addition of a helicase enzyme, crucial for initiating the unwinding of the DNA duplex by recognizing the annealed primers, action which leads to strand separation, exposing single-strand regions ready for processing ( Lee et al, 2022a , Moon et al, 2022 ). Following DNA duplex unwinding, DNA polymerase is introduced, triggering DNA synthesis by extending from the primers, utilizing single-strand DNA as templates.…”

Section: Recent Advances In Naa-based Methodsmentioning

confidence: 99%

“…As synthesis progresses, a newly formed strand displaces the complementary strand in the target DNA duplex, yielding a fresh duplex DNA structure. This cycle repeats as helicase unwinds new duplexes, primers anneal, and DNA polymerase generates strands, achieving exponential amplification of target DNA/RNA ( Lee et al, 2022a , Moon et al, 2022 ).…”

Section: Recent Advances In Naa-based Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

Ndraha,

Lin,

Wang

et al. 2023

Food Chemistry: Molecular Sciences

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Section: Recent Advances In Naa-based Methodsmentioning

confidence: 99%

Section: Recent Advances In Naa-based Methodsmentioning

confidence: 99%

Section: Recent Advances In Naa-based Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

Ndraha,

Lin,

Wang

et al. 2023

Food Chemistry: Molecular Sciences

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“…HDA has been adapted for colorimetric, fluorescent and lateral flow read-outs, making it applicable to POC and in-field applications [ 61 ]. HDA has been used to detect a large number of human and animal pathogens, including Norovirus [ 62 ], SARS-CoV-2 [ 63 ], M. tuberculosis [ 64 ], T. vaginalis [ 65 ] and yellow fever virus in a resource-limited setting [ 66 ].…”

Section: Isothermal Amplification Technologiesmentioning

confidence: 99%

Latest Advances in Arbovirus Diagnostics

2023

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Arboviruses are a diverse family of vector-borne pathogens that include members of the Flaviviridae, Togaviridae, Phenuviridae, Peribunyaviridae, Reoviridae, Asfarviridae, Rhabdoviridae, Orthomyxoviridae and Poxviridae families. It is thought that new world arboviruses such as yellow fever virus emerged in the 16th century due to the slave trade from Africa to America. Severe disease-causing viruses in humans include Japanese encephalitis virus (JEV), yellow fever virus (YFV), dengue virus (DENV), West Nile virus (WNV), Zika virus (ZIKV), Crimean–Congo hemorrhagic fever virus (CCHFV), severe fever with thrombocytopenia syndrome virus (SFTSV) and Rift Valley fever virus (RVFV). Numerous methods have been developed to detect the presence of these pathogens in clinical samples, including enzyme-linked immunosorbent assays (ELISAs), lateral flow assays (LFAs) and reverse transcriptase–polymerase chain reaction (RT-PCR). Most of these assays are performed in centralized laboratories due to the need for specialized equipment, such as PCR thermal cyclers and dedicated infrastructure. More recently, molecular methods have been developed which can be performed at a constant temperature, termed isothermal amplification, negating the need for expensive thermal cycling equipment. In most cases, isothermal amplification can now be carried out in as little as 5–20 min. These methods can potentially be used as inexpensive point of care (POC) tests and in-field deployable applications, thus decentralizing the molecular diagnosis of arboviral disease. This review focuses on the latest developments in isothermal amplification technology and detection techniques that have been applied to arboviral diagnostics and highlights future applications of these new technologies.

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“…When compared to PCR experiments, not only can both these methods be used in whole process experiments at a constant temperature, but they also have simple operation and fewer instrument and personnel requirements, making them suitable for rapid diagnosis at the grassroots or on-site level. Thus, since their development, HDA and RPA have been used extensively in medical diagnosis, agriculture, food, and biological safety, amongst other elds [10][11][12][13] . Moreover, by combining lateral ow test (LFT) and INAAT, the detection results can be observed with the naked eye.…”

Section: Introductionmentioning

confidence: 99%

Detection of Monkeypox virus Using Helicase Dependent Amplification and Recombinase Polymerase Amplification Combined with Lateral Flow Test

Gulinaizhaer,

Yang,

Zou

et al. 2023

Preprint

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The monkeypox virus (MPXV) is a zoonotic DNA virus that belongs to the poxvirus family. Conventional laboratory methods for detecting MPXV are complex and expensive, making them unsuitable for detecting the virus in regions with limited resources. In this study, we using the Helicase dependent amplification (HDA) method and the Recombinase polymerase amplification (RPA) technique in combination with the lateral flow test (LFT), together with a self-designed qPCR technique for the detection of the monkeypox virus-specific conserved fragment F3L, to compare the sensitivity and specificity of the three assays. The assay results showed that the detection limits for HDA-LFT and RPA-LFT were ༜ 10 copies/µL and for qPCR ༜ 500 copies/µL. The three methods described above showed good specificity in the detection of monkeypox viruses among their congeners. The simple, highly sensitive, and specific MPXV assay developed in this study is anticipated to provide a solid foundation for future applications in the early screening, diagnosis, and evaluation of the efficacy of MPXV. This is the first time the HDA-LFT assay has been utilized to detect MPXV infection.

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Colorimetric detection of norovirus by helicase-dependent amplification method based on specific primers integrated with HRPzyme

Cited by 8 publications

References 28 publications

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

Latest Advances in Arbovirus Diagnostics

Detection of Monkeypox virus Using Helicase Dependent Amplification and Recombinase Polymerase Amplification Combined with Lateral Flow Test

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