Nucleic Acid-based Diagnosis and Epidemiology of Infectious Diseases

Abstract Context—Molecular diagnostics continues to evolve very rapidly, and its impact in the diagnosis of infectious diseases is undeniable. Molecular tools have played a pivotal role in discovering and characterizing several emerging infectious agents and have now become the gold standard for the diagnosis of infectious diseases caused by fastidious or uncultivable agents. Multiple challenges still remain for the widespread use of cost-effective, validated, and commercially available molecular tools. Automated instruments capable of sample processing and multiplex nucleic acid amplification and postamplification analysis have already been approved by the US Food and Drug Administration (FDA) for use in the clinical setting. Nanobiotechnology is beginning to impact laboratory diagnostics in the clinical setting. Objective—To address current nucleic acid techniques used in the clinical laboratory for diagnosis of infectious diseases. FDA-approved tests are listed, as well as molecular techniques (amplification and postamplification analysis). A comprehensive list of emerging pathogens during the last 4 decades is also presented. Biosurveillance systems are discussed in the context of molecular tools. The rapidly evolving field of nanobiotechnology is briefly addressed. Data Sources—Original publications, major reviews, and book chapters were used to present a comprehensive, yet short, review of molecular diagnostics in infectious diseases. Conclusions—We will continue to witness an exponential growth of molecular techniques used for the initial diagnosis of infectious diseases. Molecular tools will also continue to have an impact on disease prognosis and response to therapeutic interventions. Automation, multiplexing, and miniaturization will continue to be driving forces in the development of new instruments.

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Current State of Commercial Point-of-Care Nucleic Acid Tests for Infectious Diseases

The Analyst ◽

10.1039/d0an01988g ◽

2021 ◽

Author(s):

Jane Zhang ◽

Andrew T. Bender ◽

David S. Boyle ◽

Paul K Drain ◽

Jonathan D. Posner

Keyword(s):

Nucleic Acid ◽

Infectious Diseases ◽

Point Of Care ◽

Current State ◽

Nucleic Acid Tests

The COVID-19 pandemic has put the spotlight on the urgent need for integrated nucleic acid tests (NATs) for infectious diseases, especially those that can be used near patient (“point-of-care”, POC),...

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Detection of potential microbial antigens by immuno-PCR (PCR-amplified immunoassay)

Journal of Medical Microbiology ◽

10.1099/jmm.0.070318-0 ◽

2014 ◽

Vol 63 (5) ◽

pp. 627-641 ◽

Cited By ~ 28

Author(s):

Promod K. Mehta ◽

Ankush Raj ◽

Netra Pal Singh ◽

Gopal K. Khuller

Keyword(s):

Nucleic Acid ◽

Infectious Diseases ◽

Real Time ◽

Magnetic Beads ◽

Fold Increase ◽

Biological Molecules ◽

Nucleic Acid Amplification ◽

Pcr Assay ◽

Immunological Research ◽

Sensitive Method

Immuno-PCR (PCR-amplified immunoassay; I-PCR) is a novel ultrasensitive method combining the versatility of ELISA with the sensitivity of nucleic acid amplification of PCR. The enormous exponential amplification power of PCR in an I-PCR assay leads to at least a 102–104-fold increase in sensitivity compared with an analogous ELISA. I-PCR has been used to detect many biological molecules such as proto-oncogenes, toxins, cytokines, hormones, and biomarkers for autoimmune and Alzheimer’s diseases, as well as microbial antigens and antibodies, and it can be adapted as a novel diagnostic tool for various infectious and non-infectious diseases. Quantitative real-time I-PCR has the potential to become the most analytically sensitive method for the detection of proteins. The sensitivity and specificity of a real-time I-PCR assay can be enhanced further with the use of magnetic beads and nanoparticles. This review is primarily focused on the detection of potential viral, bacterial and parasitic antigens by I-PCR assay, thus enabling their application for immunological research and for early diagnosis of infectious diseases.

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Rapidemic, a versatile and label-free DNAzyme-based platform for visual nucleic acid detection

10.1101/2020.10.14.337808 ◽

2020 ◽

Author(s):

Marijn van den Brink ◽

Sebastian T. Tandar ◽

Tim A. P. van den Akker ◽

Sinisha Jovikj ◽

Violette Defourt ◽

...

Keyword(s):

Nucleic Acid ◽

Infectious Diseases ◽

Detection Method ◽

Point Of Care ◽

Diagnostic Testing ◽

Nucleic Acid Detection ◽

Label Free ◽

Strand Displacement Amplification ◽

Sequence Detection ◽

Accessible Point

AbstractIn the last three decades, there have been recurring outbreaks of infectious diseases, brought to light with the recent outbreak of coronavirus disease 2019 (COVID-19). Attempts to effectively contain the spread of infectious diseases have been hampered by the lack of rapidly adaptable, accurate, and accessible point-of-care diagnostic testing. In this study, we present a novel design of a label-free DNAzyme-based detection method called Rapidemic. This assay combines recombinase polymerase amplification (RPA) with linear strand-displacement amplification (LSDA) and guanine-quadruplex (GQ) DNAzyme-catalysed colour-changing reaction. The colorimetry basis of the signal readout omits the need for extensive instrumentation. Moreover, the primer-based sequence detection of RPA gives Rapidemic a potential to be rapidly adapted to target a new sequence. As a proof of concept, we developed the assay to detect isolated genomic DNA of Saccharomyces cerevisiae. The use of low-pH buffers and the optimization of the dilution rates from each preceding reaction to the next showed to be successful strategies to enable visible detection with this method. These findings demonstrate for the first time that a label-free DNAzyme-based detection method can be coupled to RPA and LSDA for nucleic acid detection.

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The Standardization and Control of Serology and Nucleic Acid Testing for Infectious Diseases

Clinical Microbiology Reviews ◽

10.1128/cmr.00035-21 ◽

2021 ◽

Author(s):

Wayne Dimech

Keyword(s):

Infectious Disease ◽

Nucleic Acid ◽

Infectious Diseases ◽

Complement Fixation ◽

Nucleic Acid Testing ◽

Biological Functions ◽

Patient Sample ◽

Test Systems ◽

Specific Antigens ◽

And Control

Historically, the detection of antibodies against infectious disease agents was achieved using test systems that utilized biological functions such as neutralization, complement fixation, hemagglutination, or visualization of binding of antibodies to specific antigens, using testing doubling dilutions of the patient sample to determine an endpoint. These test systems have since been replaced by automated platforms, many of which have been integrated into general medical pathology.

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