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Diagnostics
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Molecular Diagnostics of Emerging Pathogens for Infectious Diseases
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Molecular Diagnostics of Emerging Pathogens for Infectious Diseases

A special issue of Diagnostics (ISSN 2075-4418). This special issue belongs to the section "Diagnostic Microbiology and Infectious Disease".

Deadline for manuscript submissions: closed (10 December 2022) | Viewed by 11787

Special Issue Editors

Dr. Melinda D. Poulter

E-Mail Website
Guest Editor
Director, Clinical Microbiology, University of Virginia, Charlottesville, VA 22901, USA
Interests: clinical microbiology; diagnostics and therapy; virology; bacteriology; mycology; molecular microbiology; parasitology; infectious disease serology
Prof. Dr. James P. Landers

E-Mail Website
Guest Editor
Departments of Chemistry, Mechanical Engineering and Pathology, Center for Nano-Bio Research, University of Virginia, Charlottesville, VA 22904-4319, USA
Interests: microfluidic systems; clinical and forensic assays; capillary electrophoresis; molecular diagnostics; microchip fabrication; “sample in–answer out” microdevices; micro-total analysis systems (microTAS)
Dr. Jamila S. Marshall

E-Mail
Guest Editor
Department of Chemistry, University of Virginia, Charlottesville, VA 22901, USA
Interests: forensic and clinical assays; microdevice fabrication; micro-total analysis systems (microTAS); microfluidic separations; bioanalytical chemistry; rapid PCR; diagnosis of infectious diseases
Dr. Rachelle A. Turiello

E-Mail
Guest Editor
Department of Chemistry, University of Virginia, Charlottesville, VA 22901, USA
Interests: epigenetics; micro-total analysis systems (microTAS); forensic and clinical assays; microdevice fabrication; rapid PCR; molecular diagnostics; DNA methylation analysis

Special Issue Information

Dear Colleagues,

In recent years, outbreaks of infectious diseases have piqued significant public and scientific interest, with the COVID-19 global pandemic being at the forefront of this charge. Emerging and reemerging pathogens, as well as the detection of consequent illnesses, have spurred a wave of exciting and

novel advances in molecular diagnostics. While the application of gold-standard techniques to these research problems addresses a large subsection of diagnostic needs, innovative, reproducible, sensitive, and specific methods for detecting pathogens are critical to the ongoing efforts to mitigate the incidence of infectious diseases in global populations. As such, the aim of this Special Issue, entitled “Molecular Diagnostics of Emerging Pathogens for Infectious Diseases”, is to elucidate advances in this field via the assessment and comparison of such diagnostic methods.

Dr. Melinda D. Poulter
Prof. Dr. James P. Landers
Dr. Jamila S. Marshall
Dr. Rachelle A. Turiello
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Diagnostics is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • infectious diseases
  • molecular diagnostics
  • pathogens
  • microbiology
  • genomics
  • biomarkers

Published Papers (7 papers)

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12 pages, 1121 KiB  
Article
SARS-CoV-2 Neutralizing Antibodies in Mexican Population: A Five Vaccine Comparison
by Fernando Alcorta-Nuñez, Diana Cristina Pérez-Ibave, Carlos Horacio Burciaga-Flores, Miguel Ángel Garza, Moisés González-Escamilla, Patricia Rodríguez-Niño, Juan Francisco González-Guerrero, Adelina Alcorta-Garza, Oscar Vidal-Gutiérrez, Genaro A. Ramírez-Correa and María Lourdes Garza-Rodríguez
Diagnostics 2023, 13(6), 1194; https://doi.org/10.3390/diagnostics13061194 - 22 Mar 2023
Cited by 3 | Viewed by 1781
Abstract
Neutralizing antibodies (NAs) are key immunological markers and are part of the humoral response of the adaptive immune system. NA assays determine the presence of functional antibodies to prevent SARS-CoV-2 infection. We performed a real-world evidence study to detect NAs that confer protection [...] Read more.
Neutralizing antibodies (NAs) are key immunological markers and are part of the humoral response of the adaptive immune system. NA assays determine the presence of functional antibodies to prevent SARS-CoV-2 infection. We performed a real-world evidence study to detect NAs that confer protection against SARS-CoV-2 after the application of five vaccines (Pfizer/BioNTech, AstraZeneca, Sinovac, Moderna, and CanSino) in the Mexican population. Side effects of COVID-19 vaccines and clinical and demographic factors associated with low immunogenicity were also evaluated. A total of 242 SARS-CoV-2-vaccinated subjects were recruited. Pfizer/BioNTech and Moderna proved the highest percentage of inhibition in a mono-vaccine scheme. Muscular pain, headache, and fatigue were the most common adverse events. None of the patients reported severe adverse events. We found an estimated contagion-free time of 207 (IQR: 182–231) and 187 (IQR: 184–189) days for Pfizer/BioNTech and CanSino in 12 cases in each group. On the basis of our results, we consider that the emerging vaccination strategy in Mexico is effective and safe. Full article
(This article belongs to the Special Issue Molecular Diagnostics of Emerging Pathogens for Infectious Diseases)
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7 pages, 723 KiB  
Communication
Evaluation of Crude and Recombinant Antigens of Schistosoma japonicum for the Detection of Schistosoma mekongi Human Infection
by Jose Ma. M. Angeles, Atcharaphan Wanlop, Minh-Anh Dang-Trinh, Masashi Kirinoki, Shin-ichiro Kawazu and Aya Yajima
Diagnostics 2023, 13(2), 184; https://doi.org/10.3390/diagnostics13020184 - 04 Jan 2023
Cited by 1 | Viewed by 1510
Abstract
Asian schistosomiasis caused by the blood fluke Schistosoma mekongi is endemic in northern Cambodia and Southern Lao People’s Democratic Republic. The disease is mainly diagnosed by stool microscopy. However, serodiagnosis such as enzyme-linked immunosorbent assay (ELISA) with soluble egg antigen (SEA), has been [...] Read more.
Asian schistosomiasis caused by the blood fluke Schistosoma mekongi is endemic in northern Cambodia and Southern Lao People’s Democratic Republic. The disease is mainly diagnosed by stool microscopy. However, serodiagnosis such as enzyme-linked immunosorbent assay (ELISA) with soluble egg antigen (SEA), has been shown to have better sensitivity compared to the stool examination, especially in the settings with a low intensity of infection. To date, no recombinant antigen has been assessed using ELISA for the detection of S. mekongi infection, due to the lack of genome information for this schistosome species. Thus, the objective of this study is to evaluate several recombinant S. japonicum antigens that have been developed in our laboratory for the detection of S. mekongi infection. The crude antigen SjSEA and recombinant antigens Sj7TR, SjPCS, SjPRx-4, and SjChi-3 were evaluated in ELISA using serum samples positive for S. mekongi infection. The cross-reaction was checked using sera positive for Ophistorchis viverrini. ELISA results showed that S. japonicum SEA at low concentrations showed better diagnostic performance than the recombinant antigens tested using the archived serum samples from Cambodia. However, further optimization of the recombinant antigens should be conducted in future studies to improve their diagnostic performance for S. mekongi detection. Full article
(This article belongs to the Special Issue Molecular Diagnostics of Emerging Pathogens for Infectious Diseases)
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9 pages, 967 KiB  
Article
Cloning, Expression and Evaluation of Thioredoxin Peroxidase-1 Antigen for the Serological Diagnosis of Schistosoma mekongi Human Infection
by Atcharaphan Wanlop, Jose Ma. M. Angeles, Adrian Miki C. Macalanda, Masashi Kirinoki, Yuma Ohari, Aya Yajima, Junya Yamagishi, Kevin Austin L. Ona and Shin-ichiro Kawazu
Diagnostics 2022, 12(12), 3077; https://doi.org/10.3390/diagnostics12123077 - 07 Dec 2022
Viewed by 1425
Abstract
Schistosoma mekongi, a blood fluke that causes Asian zoonotic schistosomiasis, is distributed in communities along the Mekong River in Cambodia and Lao People’s Democratic Republic. Decades of employing numerous control measures including mass drug administration using praziquantel have resulted in a decline [...] Read more.
Schistosoma mekongi, a blood fluke that causes Asian zoonotic schistosomiasis, is distributed in communities along the Mekong River in Cambodia and Lao People’s Democratic Republic. Decades of employing numerous control measures including mass drug administration using praziquantel have resulted in a decline in the prevalence of schistosomiasis mekongi. This, however, led to a decrease in sensitivity of Kato–Katz stool microscopy considered as the gold standard in diagnosis. In order to develop a serological assay with high sensitivity and specificity which can replace Kato–Katz, recombinant S. mekongi thioredoxin peroxidase-1 protein (rSmekTPx-1) was expressed and produced. Diagnostic performance of the rSmekTPx-1 antigen through ELISA for detecting human schistosomiasis was compared with that of recombinant protein of S. japonicum TPx-1 (rSjTPx-1) using serum samples collected from endemic foci in Cambodia. The sensitivity and specificity of rSmekTPx-1 in ELISA were 89.3% and 93.3%, respectively, while those of rSjTPx-1 were 71.4% and 66.7%, respectively. In addition, a higher Kappa value of 0.82 calculated between rSmekTPx-1 antigen ELISA and Kato–Katz confirmed better agreement than between rSjTPx-1 antigen ELISA and Kato–Katz (Kappa value 0.38). These results suggest that ELISA with rSmekTPx-1 antigen can be a potential diagnostic method for detecting active human S. mekongi infection. Full article
(This article belongs to the Special Issue Molecular Diagnostics of Emerging Pathogens for Infectious Diseases)
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10 pages, 2239 KiB  
Article
Improved Detection of Herpesviruses from Diluted Vitreous Specimens Using Hydrogel Particles
by Nicole L. Belanger, Robbie Barbero, Robert Barclay, Benjamin Lepene, Lucia Sobrin and Paulo J. M. Bispo
Diagnostics 2022, 12(12), 3016; https://doi.org/10.3390/diagnostics12123016 - 01 Dec 2022
Viewed by 1136
Abstract
Infectious uveitis is a sight-threatening infection commonly caused by herpesviruses. Vitreous humor is often collected for molecular confirmation of the causative agent during vitrectomy and mixed in large volumes of buffered saline, diluting the pathogen load. Here, we explore affinity-capture hydrogel particles (Nanotrap [...] Read more.
Infectious uveitis is a sight-threatening infection commonly caused by herpesviruses. Vitreous humor is often collected for molecular confirmation of the causative agent during vitrectomy and mixed in large volumes of buffered saline, diluting the pathogen load. Here, we explore affinity-capture hydrogel particles (Nanotrap®) to concentrate low abundant herpesviruses from diluted vitreous. Simulated samples were prepared using porcine vitreous spiked with HSV-1, HSV-2, VZV and CMV at 105 copies/mL. Pure undiluted samples were used to test capturing capability of three custom Nanotrap particles (red, white and blue) in a vitreous matrix. We found that all particles demonstrated affinity to the herpesviruses, with the Red Particles having both good capture capability and ease of handling for all herpesviruses. To mimic diluted vitrectomy specimens, simulated-infected vitreous were then serially diluted in 7 mL TE buffer. Diluted samples were subjected to an enrichment protocol using the Nanotrap Red particles. Sensitivity of pathogen detection by qPCR in diluted vitreous increased anywhere between 2.3 to 26.5 times compared to non-enriched specimens. This resulted in a 10-fold increase in the limit of detection for HSV-1, HSV-2 and VZV. These data demonstrated that Nanotrap particles can capture and concentrate HSV-1, HSV-2, VZV and CMV in a vitreous matrix. Full article
(This article belongs to the Special Issue Molecular Diagnostics of Emerging Pathogens for Infectious Diseases)
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9 pages, 1623 KiB  
Article
Calprotectin Levels and Neutrophil Count Are Prognostic Markers of Mortality in COVID-19 Patients
by Giovanna Cardiero, Daniela Palma, Martina Vano, Claudia Anastasio, Biagio Pinchera, Martina Ferrandino, Carlo Gianfico, Luca Gentile, Marcella Savoia, Ivan Gentile, Maria Donata Di Taranto and Giuliana Fortunato
Diagnostics 2022, 12(10), 2554; https://doi.org/10.3390/diagnostics12102554 - 20 Oct 2022
Cited by 5 | Viewed by 1475
Abstract
Inflammation plays a crucial role in worsening coronavirus disease (COVID-19). Calprotectin is a pro-inflammatory molecule produced by monocytes and neutrophilic granulocytes. The aim of the study was to evaluate both the prognostic role of circulating calprotectin levels and neutrophil count toward fatal outcome [...] Read more.
Inflammation plays a crucial role in worsening coronavirus disease (COVID-19). Calprotectin is a pro-inflammatory molecule produced by monocytes and neutrophilic granulocytes. The aim of the study was to evaluate both the prognostic role of circulating calprotectin levels and neutrophil count toward fatal outcome in COVID-19 patients. We retrospectively collected and analyzed data on 195 COVID-19 adult patients, 156 hospitalized in the infectious disease unit and 39 in the intensive care unit (ICU). Calprotectin levels and neutrophil counts measured at the first hospitalization day were higher in the patients with a fatal outcome than in surviving ones. The association of high calprotectin levels and neutrophil count to patient death remain significant by logistic regression, independent of patient age. ROC curves analysis for calprotectin levels and neutrophil count revealed a good discriminatory power toward survival (area under the curve of 0.759 and 0.843, respectively) and identified the best cut-off (1.66 mg/L and 16.39 × 103/µL, respectively). Kaplan–Meier analysis confirmed the prognostic role of high calprotectin levels and neutrophil count in death prediction. In conclusion, this study highlights that calprotectin levels together with neutrophil count should be considered as biomarkers of mortality in COVID-19 patients. Full article
(This article belongs to the Special Issue Molecular Diagnostics of Emerging Pathogens for Infectious Diseases)
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15 pages, 2172 KiB  
Article
Rapid SARS-CoV-2 Virus Enrichment and RNA Extraction for Efficient Diagnostic Screening of Pooled Nasopharyngeal or Saliva Samples for Dilutions Up to 1:100
by Jamila S. Marshall, Rachelle Turiello, Larissa L. Cunha, Ella V. Frazier, Jeff Hickey, Jeff Chapman, Melinda D. Poulter, Heather L. Fehling and James P. Landers
Diagnostics 2022, 12(6), 1398; https://doi.org/10.3390/diagnostics12061398 - 06 Jun 2022
Cited by 1 | Viewed by 2344
Abstract
As COVID-19 transmission control measures are gradually being lifted, a sensitive and rapid diagnostic method for large-scale screening could prove essential for monitoring population infection rates. However, many rapid workflows for SARS-CoV-2 detection and diagnosis are not amenable to the analysis of large-volume [...] Read more.
As COVID-19 transmission control measures are gradually being lifted, a sensitive and rapid diagnostic method for large-scale screening could prove essential for monitoring population infection rates. However, many rapid workflows for SARS-CoV-2 detection and diagnosis are not amenable to the analysis of large-volume samples. Previously, our group demonstrated a technique for SARS-CoV-2 nanoparticle-facilitated enrichment and enzymatic lysis from clinical samples in under 10 min. Here, this sample preparation strategy was applied to pooled samples originating from nasopharyngeal (NP) swabs eluted in viral transport medium (VTM) and saliva samples diluted up to 1:100. This preparation method was coupled with conventional RT-PCR on gold-standard instrumentation for proof-of-concept. Additionally, real-time PCR analysis was conducted using an in-house, ultra-rapid real-time microfluidic instrument paired with an experimentally optimized rapid protocol. Following pooling and extraction from clinical samples, average cycle threshold (CT) values from resultant eluates generally increased as the pooling dilution factor increased; further, results from a double-blind study demonstrated 100% concordance with clinical values. In addition, preliminary data obtained from amplification of eluates prepared by this technique and analyzed using our portable, ultra-rapid real-time microfluidic PCR amplification instrument showed progress toward a streamlined method for rapid SARS-CoV-2 analysis from pooled samples. Full article
(This article belongs to the Special Issue Molecular Diagnostics of Emerging Pathogens for Infectious Diseases)
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7 pages, 525 KiB  
Case Report
Tularemia Presenting Solely with Cervical Lymphadenopathy and Fever
by Göran Ramin Boeckel, Jan Basri Adiprasito, Neele Judith Froböse, Frieder Schaumburg, Richard Vollenberg and Phil-Robin Tepasse
Diagnostics 2022, 12(8), 2000; https://doi.org/10.3390/diagnostics12082000 - 18 Aug 2022
Cited by 1 | Viewed by 1427
Abstract
A 52-year-old German female presented with cervical lymphadenopathy and fever. Despite the initial symptom-presentation leading to the consideration of sarcoidosis, lymphoma, tuberculosis, and toxoplasmosis, an extensive serologic and histo- and molecular pathologic workup eventually indicated a likely diagnosis of tularemia. This case brings [...] Read more.
A 52-year-old German female presented with cervical lymphadenopathy and fever. Despite the initial symptom-presentation leading to the consideration of sarcoidosis, lymphoma, tuberculosis, and toxoplasmosis, an extensive serologic and histo- and molecular pathologic workup eventually indicated a likely diagnosis of tularemia. This case brings to light that tularemia is a diagnostic challenge and requires high reliance on the epidemiological context thorough patient history, and an extensive interdisciplinary diagnostic workup. Full article
(This article belongs to the Special Issue Molecular Diagnostics of Emerging Pathogens for Infectious Diseases)
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