Diagnosing Emerging and Reemerging Infectious Diseases: The Pivotal Role of the Pathologist

2011 ◽  
Vol 135 (1) ◽  
pp. 83-91 ◽  
Author(s):  
Juan P. Olano ◽  
David H. Walker
Keyword(s):  
Nucleic Acid ◽  
Infectious Diseases ◽  
Molecular Diagnostics ◽  
Clinical Setting ◽  
Driving Forces ◽  
Molecular Techniques ◽  
Nucleic Acid Amplification ◽  
Pivotal Role ◽  
Laboratory Diagnostics ◽  
Molecular Tools

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.

scholarly journals Universal amplification-free molecular diagnostics by billion-fold hierarchical nanofluidic concentration

2019 ◽  
Vol 116 (33) ◽  
pp. 16240-16249 ◽  
Author(s):  
Wei Ouyang ◽  
Jongyoon Han
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Molecular Diagnostics ◽  
Troponin I ◽  
Direct Detection ◽  
Signal To Noise Ratio ◽  
Protein Detection ◽  
Clinical Diagnostics ◽  
Nucleic Acid Amplification ◽  
Enzyme Linked Immunosorbent Assay

Rapid and reliable detection of ultralow-abundance nucleic acids and proteins in complex biological media may greatly advance clinical diagnostics and biotechnology development. Currently, nucleic acid tests rely on enzymatic processes for target amplification (e.g., PCR), which have many inherent issues restricting their implementation in diagnostics. On the other hand, there exist no protein amplification techniques, greatly limiting the development of protein-based diagnosis. We report a universal biomolecule enrichment technique termed hierarchical nanofluidic molecular enrichment system (HOLMES) for amplification-free molecular diagnostics using massively paralleled and hierarchically cascaded nanofluidic concentrators. HOLMES achieves billion-fold enrichment of both nucleic acids and proteins within 30 min, which not only overcomes many inherent issues of nucleic acid amplification but also provides unprecedented enrichment performance for protein analysis. HOLMES features the ability to selectively enrich target biomolecules and simultaneously deplete nontargets directly in complex crude samples, thereby enormously enhancing the signal-to-noise ratio of detection. We demonstrate the direct detection of attomolar nucleic acids in urine and serum within 35 min and HIV p24 protein in serum within 60 min. The performance of HOLMES is comparable to that of nucleic acid amplification tests and near million-fold improvement over standard enzyme-linked immunosorbent assay (ELISA) for protein detection, being much simpler and faster in both applications. We additionally measured human cardiac troponin I protein in 9 human plasma samples, and showed excellent agreement with ELISA and detection below the limit of ELISA. HOLMES is in an unparalleled position to unleash the potential of protein-based diagnosis.

Emerging ultrafast nucleic acid amplification technologies for next-generation molecular diagnostics

2019 ◽  
Vol 141 ◽  
pp. 111448 ◽  
Author(s):  
Sang Hun Lee ◽  
Seung-min Park ◽  
Brian N. Kim ◽  
Oh Seok Kwon ◽  
Won-Yep Rho ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnostics ◽  
Nucleic Acid Amplification ◽  
Next Generation

scholarly journals CURRENT STATE OF THE PROBLEM OF THE LABORATORY DIAGNOSTICS OF UROGENITAL TRICHOMONIASIS

2013 ◽  
Vol 62 (1) ◽  
pp. 32-41
Author(s):  
Aleksey Nikolayevich Grigoryev
Keyword(s):  
Nucleic Acid ◽  
Reproductive Tract ◽  
Review Literature ◽  
Nucleic Acid Amplification ◽  
Immunological Methods ◽  
Laboratory Diagnostics ◽  
Actual Problem ◽  
Culture Techniques ◽  
Current State ◽  
Wet Mount

The laboratory diagnostics of urogenital trichomoniasis is an actual problem of modern microbiology of infections of the reproductive tract. In the review, literature data on methods of microbiological diagnostics of trichomoniasis are presented, which include microscopy of wet mount and stained preparations, culture techniques, immunological methods and nucleic acid amplification tests.

Translating Nucleic Acid Amplification Assays to the Microscale: Lab on a Chip for Point-of-Care Molecular Diagnostics

2016 ◽  
Vol 12 (5) ◽  
pp. 386-396 ◽  
Author(s):  
Michael G. Mauk ◽  
Jinzhao Song ◽  
Yubing Tong ◽  
Haim H. Bau ◽  
Changchun Liu
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnostics ◽  
Point Of Care ◽  
Lab On A Chip ◽  
Nucleic Acid Amplification

scholarly journals fM to aM nucleic acid amplification for molecular diagnostics in a non-stick-coated metal microfluidic bioreactor

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Guoliang Huang ◽  
Qin Huang ◽  
Li Ma ◽  
Xianbo Luo ◽  
Biao Pang ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnostics ◽  
Nucleic Acid Amplification ◽  
Coated Metal

scholarly journals A Simple, Inexpensive Device for Nucleic Acid Amplification without Electricity—Toward Instrument-Free Molecular Diagnostics in Low-Resource Settings

PLoS ONE ◽  
2011 ◽  
Vol 6 (5) ◽  
pp. e19738 ◽  
Author(s):  
Paul LaBarre ◽  
Kenneth R. Hawkins ◽  
Jay Gerlach ◽  
Jared Wilmoth ◽  
Andrew Beddoe ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnostics ◽  
Nucleic Acid Amplification ◽  
Low Resource Settings ◽  
Low Resource

scholarly journals Molecular diagnostics in a teacup: Non-Instrumented Nucleic Acid Amplification (NINA) for rapid, low cost detection of Salmonella enterica

2013 ◽  
Vol 58 (10) ◽  
pp. 1162-1168 ◽  
Author(s):  
Ryo Kubota ◽  
Paul Labarre ◽  
Bernhard H. Weigl ◽  
Yong Li ◽  
Paul Haydock ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnostics ◽  
Salmonella Enterica ◽  
Low Cost ◽  
Nucleic Acid Amplification

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

2014 ◽  
Vol 63 (5) ◽  
pp. 627-641 ◽  
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.

scholarly journals Comparison of Antigen Tests and qPCR in Rapid Diagnostics of Infections Caused by SARS-CoV-2 Virus

Viruses ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 17
Author(s):  
Adrianna Klajmon ◽  
Aldona Olechowska-Jarząb ◽  
Dominika Salamon ◽  
Agnieszka Sroka-Oleksiak ◽  
Monika Brzychczy-Włoch ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Molecular Techniques ◽  
Nucleic Acid Amplification ◽  
Laboratory Diagnostics ◽  
Test Results ◽  
Rapid Diagnostics ◽  
Antigen Test ◽  
Qualified Personnel ◽  
Replication Phase ◽  
Antigen Tests

Diagnostics of the coronavirus disease 2019 (COVID-19) using molecular techniques from the collected respiratory swab specimens requires well-equipped laboratory and qualified personnel, also it needs several hours of waiting for results and is expensive. Antigen tests appear to be faster and cheaper but their sensitivity and specificity are debatable. The aim of this study was to compare a selected antigen test with quantitative polymerase chain reaction (qPCR) tests results. Nasopharyngeal swabs were collected from 192 patients with COVID-19 symptoms. All samples were tested using Vitassay qPCR SARS-CoV-2 kit and the Humasis COVID-19 Ag Test (MedSun) antigen immunochromatographic test simultaneously. Ultimately, 189 samples were tested; 3 samples were excluded due to errors in taking swabs. The qPCR and antigen test results were as follows: 47 positive and 142 negative, and 45 positive and 144 negative, respectively. Calculated sensitivity of 91.5% and specificity of 98.6% for the antigen test shows differences which are not statistically significant in comparison to qPCR. Our study showed that effectiveness of the antigen tests in rapid laboratory diagnostics is high enough to be an alternative and support for nucleic acid amplification tests (NAAT) in the virus replication phase in the course of COVID-19.

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