Simple and feasible detection of hepatitis a virus using reverse transcription multienzyme isothermal rapid amplification and lateral flow dipsticks without standard PCR laboratory

Sun, Mao-Ling; Zhong, Yuan; Li, Xiaona; Yao, Jun; Pan, Yu‐Qing

doi:10.1080/21691401.2023.2203198

Cited by 6 publications

(6 citation statements)

References 44 publications

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“…All the templates used in specificity analysis were 5×10 8 plasmid DNA contains the target virus sequence. Additionally, we assessed the MIRA-LFD reaction’s specificity using the human genome as the template ( Sun et al., 2023 ).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, a few feasible and rapid isothermal nucleic detection techniques have been established. Multienzyme isothermal recombinase amplification (MIRA) is a revolutionary nucleic acid amplification technology which is based on recombinase polymerase amplification (RPA) ( Sun et al., 2023 ). Recombinase-Rec A, DNA helicase-gp41, single-stranded binding (SSB) protein, and DNA polymerase I are the four main proteins used in the procedure ( Piepenburg et al., 2006 ).…”

Section: Introductionmentioning

confidence: 99%

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Rapid detection of human cytomegalovirus by multienzyme isothermal rapid amplification and lateral flow dipsticks

Liu,

Guo,

Sun

et al. 2024

Front. Cell. Infect. Microbiol.

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IntroductionHuman cytomegalovirus (HCMV) is the most common viral infection seen in newborns. The major route of transmission for acquired human cytomegalovirus infection is breast milk from mothers who are HCMV seropositive to the infants. Thus, a rapid, economical, and simple method to perform HCMV test in breast milk is crucial and necessary for preventing acquired HCMV infection, especially in underdeveloped regions with limited laboratory resources.MethodsIn this study, an effective technique for the detection of HCMV was constructed by combining multienzyme isothermal rapid amplification (MIRA) and lateral flow chromatography strip (LFD). Primers for the conserved HCMV sequence UL83 were utilized for MIRA-LFD testing.ResultsOur results showed that the entire MIRA reaction could be completed in 12 minutes at 37°C, and LFD outcomes could be observed visibly after 10 minutes. The detection sensitivity of this method reached 50 copy/μl. Samples of breast milk were examined to compare MIRA-LFD and conventional qPCR. The accuracy of MIRA-LFD was 100%.DiscussionThe straightforward, rapid, economic features of the test can provide the significant advantages for the prevention of breast milk-acquired cytomegalovirus infection, particularly in resource-limited locations with high seroprevalence of cytomegalovirus.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid detection of human cytomegalovirus by multienzyme isothermal rapid amplification and lateral flow dipsticks

Liu,

Guo,

Sun

et al. 2024

Front. Cell. Infect. Microbiol.

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“…However, this assay uses laboratory approaches for extracting and purifying HAV RNA from stool samples. An RPA assay that is capable of detecting HAV RNA from 0.5 mL of whole blood treated with a rapid nucleic acid extraction reagent in 30 min using lateral flow strips for visual interpretation of results was reported to have perfect sensitivity and specificity when evaluated using a small number of samples [72]. These examples suggest that a POC test for the detection of HAV RNA may be possible; however, additional developments and evaluations, especially related to using sample types and sample processing methods that are relevant to a POC format are needed.…”

Section: Hepatitis a Virusmentioning

confidence: 99%

Point-of-Care Testing for Hepatitis Viruses: A Growing Need

Pauly,

Ganova-Raeva

2023

Life

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Viral hepatitis, caused by hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), or hepatitis E virus (HEV), is a major global public health problem. These viruses cause millions of infections each year, and chronic infections with HBV, HCV, or HDV can lead to severe liver complications; however, they are underdiagnosed. Achieving the World Health Organization’s viral hepatitis elimination goals by 2030 will require access to simpler, faster, and less expensive diagnostics. The development and implementation of point-of-care (POC) testing methods that can be performed outside of a laboratory for the diagnosis of viral hepatitis infections is a promising approach to facilitate and expedite WHO’s elimination targets. While a few markers of viral hepatitis are already available in POC formats, tests for additional markers or using novel technologies need to be developed and validated for clinical use. Potential methods and uses for the POC testing of antibodies, antigens, and nucleic acids that relate to the diagnosis, monitoring, or surveillance of viral hepatitis infections are discussed here. Unmet needs and areas where additional research is needed are also described.

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“…The use of helicase (gp41) and recombinases ( Streptomyces coelicolor recA, SC-recA) allows for rapid amplification that can happen between 10–30 min at a constant temperature between 35–42 °C [ 68 , 69 ]. MIRA has also been demonstrated with paper strips and paper microfluidic platforms [ 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 ]. Like the RAA, all paper-based MIRA assays utilized LFIA with colorimetric detection.…”

Section: Raa and Mira With Paper Microfluidicsmentioning

confidence: 99%

Recent Uses of Paper Microfluidics in Isothermal Nucleic Acid Amplification Tests

Reynolds,

Loeffler,

Leigh

et al. 2023

Biosensors

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Isothermal nucleic acid amplification tests have recently gained popularity over polymerase chain reaction (PCR), as they only require a constant temperature and significantly simplify nucleic acid amplification. Recently, numerous attempts have been made to incorporate paper microfluidics into these isothermal amplification tests. Paper microfluidics (including lateral flow strips) have been used to extract nucleic acids, amplify the target gene, and detect amplified products, all toward automating the process. We investigated the literature from 2020 to the present, i.e., since the onset of the COVID-19 pandemic, during which a significant surge in isothermal amplification tests has been observed. Paper microfluidic detection has been used extensively for recombinase polymerase amplification (RPA) and its related methods, along with loop-mediated isothermal amplification (LAMP) and rolling circle amplification (RCA). Detection was conducted primarily with colorimetric and fluorometric methods, although a few publications demonstrated flow distance- and surface-enhanced Raman spectroscopic (SERS)-based detection. A good number of publications could be found that demonstrated both amplification and detection on paper microfluidic platforms. A small number of publications could be found that showed extraction or all three procedures (i.e., fully integrated systems) on paper microfluidic platforms, necessitating the need for future work.

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Simple and feasible detection of hepatitis a virus using reverse transcription multienzyme isothermal rapid amplification and lateral flow dipsticks without standard PCR laboratory

Cited by 6 publications

References 44 publications

Rapid detection of human cytomegalovirus by multienzyme isothermal rapid amplification and lateral flow dipsticks

Rapid detection of human cytomegalovirus by multienzyme isothermal rapid amplification and lateral flow dipsticks

Point-of-Care Testing for Hepatitis Viruses: A Growing Need

Recent Uses of Paper Microfluidics in Isothermal Nucleic Acid Amplification Tests

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