Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay

Zasada, Aleksandra Anna; Mosiej, Ewa; Prygiel, Marta; Polak, Maciej; Wdowiak, Karol; Formińska, Kamila; Ziółkowski, Robert; Żukowski, Kamil; Marchlewicz, Kasper; Nowiński, Adam; Nowińska, Julia; Rastawicki, Waldemar; Malinowska, Elżbieta

doi:10.3390/biomedicines10092329

Cited by 23 publications

(14 citation statements)

References 37 publications

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“…HDA assays for Chlamydia trachomatis, Neisseria gonorrheae and Trichomonas vaginalis were developed by the group of Klapperich. [33][34][35] The application of two nucleic acid isothermal amplification methods-reverse transcription helicase dependent amplification (RT-HDA) and reverse transcription loop-mediated amplification (RT-LAMP), combined with a lateral flow assay, for the rapid detection of SARS_CoV_2, was presented by Zasada et al 36 (Fig. 3).…”

Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Point-of-care nucleic acid tests: assays and devices

2023

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Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Point-of-care nucleic acid tests: assays and devices

2023

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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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Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay

Cited by 23 publications

References 37 publications

Point-of-care nucleic acid tests: assays and devices

Point-of-care nucleic acid tests: assays and devices

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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