Nucleic Acids Analytical Methods for Viral Infection Diagnosis: State-of-the-Art and Future Perspectives

Sciuto, Emanuele Luigi; Leonardi, Antonio Alessio; Calabrese, Giovanna; Luca, Giovanna De; Coniglio, Maria Anna; Irrera, Alessia; Conoci, Sabrina

doi:10.3390/biom11111585

Cited by 11 publications

(10 citation statements)

References 67 publications

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“…Although NATs are frequently used in the diagnosis of infectious diseases, due to the time, cost, and effort involved and the need for expensive machines, they have begun to be replaced by time-saving, portable, and more integrated technologies named point of care (POC). This was particularly true during the COVID-19 pandemic [ 11 ].…”

Section: Current Molecular Methods For the Identification Of Viral In...mentioning

confidence: 99%

Managing Viral Emerging Infectious Diseases via Current and Future Molecular Diagnostics

Altındiş

KILBAŞ

2023

Diagnostics

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Emerging viral infectious diseases have been a constant threat to global public health in recent times. In managing these diseases, molecular diagnostics has played a critical role. Molecular diagnostics involves the use of various technologies to detect the genetic material of various pathogens, including viruses, in clinical samples. One of the most commonly used molecular diagnostics technologies for detecting viruses is polymerase chain reaction (PCR). PCR amplifies specific regions of the viral genetic material in a sample, making it easier to detect and identify viruses. PCR is particularly useful for detecting viruses that are present in low concentrations in clinical samples, such as blood or saliva. Another technology that is becoming increasingly popular for viral diagnostics is next-generation sequencing (NGS). NGS can sequence the entire genome of a virus present in a clinical sample, providing a wealth of information about the virus, including its genetic makeup, virulence factors, and potential to cause an outbreak. NGS can also help identify mutations and discover new pathogens that could affect the efficacy of antiviral drugs and vaccines. In addition to PCR and NGS, there are other molecular diagnostics technologies that are being developed to manage emerging viral infectious diseases. One of these is CRISPR-Cas, a genome editing technology that can be used to detect and cut specific regions of viral genetic material. CRISPR-Cas can be used to develop highly specific and sensitive viral diagnostic tests, as well as to develop new antiviral therapies. In conclusion, molecular diagnostics tools are critical for managing emerging viral infectious diseases. PCR and NGS are currently the most commonly used technologies for viral diagnostics, but new technologies such as CRISPR-Cas are emerging. These technologies can help identify viral outbreaks early, track the spread of viruses, and develop effective antiviral therapies and vaccines.

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Section: Current Molecular Methods For the Identification Of Viral In...mentioning

confidence: 99%

Managing Viral Emerging Infectious Diseases via Current and Future Molecular Diagnostics

Altındiş

KILBAŞ

2023

Diagnostics

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“…However, PCR-based methods need to be performed by trained personnel and in specialized laboratories and also are affected by expensive reagents and long procedures due to thermal cycling of amplification. [33][34][35] Loop-mediated isothermal amplification (LAMP) methods have been introduced to contain these limitations. LAMP are performed at constant temperature, excluding the thermal cycling typical of the PCR reaction and simplifying the analysis.…”

Section: Sars-cov-2 Conventional Analysismentioning

confidence: 99%

“…sample. [33] Most of the available molecular methods for SARS-CoV-2 are based on the digital droplet PCR (ddPCR) achieving a high sensitivity, down to 10 −2 copies mL −1 of viral genome. [40,41] The conventional methods above described do not fulfill the need to merge a sensitive quantification at the molecular level (10 copies of virus) together with rapid detection (within 15 min).…”

Section: Sars-cov-2 Conventional Analysismentioning

confidence: 99%

PCR‐Free Innovative Strategies for SARS‐CoV‐2 Detection

Calorenni,

Leonardi,

Sciuto

et al. 2023

Adv Healthcare Materials

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The pandemic outbreak caused by SARS‐CoV‐2 coronavirus brought a crucial issue in public health causing up to now more than 600 million infected people and 6.5 million deaths. Conventional diagnostic methods are based on quantitative reverse transcription polymerase chain reaction (RT‐qPCR assay) and immuno‐detection (ELISA assay). However, despite these techniques have the advantages of being standardized and consolidated, they keep some main limitations in terms of accuracy (immunoassays), time/cost consumption of analysis, the need for qualified personnel, and lab constrain (molecular assays). There is crucial the need to develop new diagnostic approaches for accurate, fast and portable viral detection and quantification. Among these, PCR‐free biosensors represent the most appealing solution since they can allow molecular detection without the complexity of the PCR. This will enable the possibility to be integrated in portable and low‐cost systems for massive and decentralized screening of SARS‐CoV‐2 in a point‐of‐care (PoC) format, pointing to achieve a performant identification and control of infection. In this review, the most recent approaches for the SARS‐CoV‐2 PCR‐free detection are reported, describing both the instrumental and methodological features, and highlighting their suitability for a PoC application.

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“…In contrast to immunoassays, nucleic acid detection has several distinct advantages, including the high specificity of recognition, the relative ease of developing new oligonucleotide primers for new assays and/or novel pathogens, and the inherent potential for exponential amplification arising from the nature of the replicative mechanism. PCR is the gold-standard for nucleic acid amplification, and great progress has been made in adapting it for point-of-use applications [ 7 ], including with electrochemical sensors [ 8 ]. Despite these advances, however, PCR still has a relatively long time-to-detection, and the requirement for thermocycling increases device complexity.…”

Section: Introductionmentioning

confidence: 99%

Strategies for the Voltammetric Detection of Loop-Mediated Isothermal Amplification

Marangoni

Emadi

2023

Micromachines

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Loop-mediated isothermal amplification (LAMP) is rapidly developing into an important tool for the point-of-use detection of pathogens for both clinical and environmental samples, largely due to its sensitivity, rapidity, and adaptability to portable devices. Many methods are used to monitor LAMP, but not all are amenable to point-of-use applications. Common methods such as fluorescence often require bulky equipment, whereas colorimetric and turbidimetric methods can lack sensitivity. Electrochemical biosensors are becoming increasingly important for these applications due to their potential for low cost, high sensitivity, and capacity for miniaturization into integrated devices. This review provides an overview of the use of voltammetric sensors for monitoring LAMP, with a specific focus on how electroactive species are used to interface between the biochemical products of the LAMP reaction and the voltammetric sensor. Various strategies for the voltammetric detection of DNA amplicons as well as pyrophosphate and protons released during LAMP are presented, ranging from direct DNA binding by electroactive species to the creative use of pyrophosphate-detecting aptamers and pH-sensitive oligonucleotide structures. Hurdles for adapting these devices to point-of-use applications are also discussed.

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Nucleic Acids Analytical Methods for Viral Infection Diagnosis: State-of-the-Art and Future Perspectives

Cited by 11 publications

References 67 publications

Managing Viral Emerging Infectious Diseases via Current and Future Molecular Diagnostics

Managing Viral Emerging Infectious Diseases via Current and Future Molecular Diagnostics

PCR‐Free Innovative Strategies for SARS‐CoV‐2 Detection

Strategies for the Voltammetric Detection of Loop-Mediated Isothermal Amplification

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