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Novel Approaches for Diagnosis of Parasitic Disease

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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 (25 October 2022) | Viewed by 12465

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Special Issue Editor

Prof. Dr. Franco H. Falcone

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Guest Editor

Institute for Parasitology, Faculty of Veterinary Medicine, Justus Liebig University Giessen, 35392 Giessen, Germany
Interests: mechanisms of Helicobacter pylori adhesion to the human gastric epithelium; development of new reporter systems for detection of sensitisation with allergen-specific IgE; molecular immunology
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Dear Colleagues,

Parasitic diseases occur worldwide and their impact on global health is very large. Commercial technologies for use in the diagnosis of common diseases such as allergic sensitization, microbial or viral diseases range from diagnostic kits to semi-automated or fully automated technologies.

In contrast, diagnostic techniques for parasitic infections, in particular for so-called Neglected Tropical Diseases (NTDs), are frequently seen in the form of ‘self-made’, internally validated diagnostic techniques in various labs. In the context of parasitic diseases, and in particular in parasitic worm infections, one of the complications in serological diagnosis is the very high level of cross-reactivity between related parasites at the genus, family or higher taxonomic level.

The aim of the Special Issue, entitled "Novel Approaches for Diagnosis of Parasitic Disease", is to explore the status quo of novel approaches with a potential to overcome the impasse in the area of parasitic disease diagnosis.

Prof. Dr. Franco Falcone
Guest Editor

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

arasites
Diagnosis
Neglected Tropical Diseases (NTDs)
Array technologies
Multiplex technologies
Microfluidics
Reporter assays.

Published Papers (6 papers)

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Research

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12 pages, 9847 KiB

Open AccessArticle

Polyclonal Antibody Generation against PvTRAg for the Development of a Diagnostic Assay for Plasmodium vivax

by Shalini Aggarwal, Selvamano Selvaraj, Jayaprakash Nattamai Subramanian, Mookambeswaran Arunachalam Vijayalakshmi, Swati Patankar and Sanjeeva Srivastava

Diagnostics 2023, 13(5), 835; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics13050835 - 22 Feb 2023

Viewed by 1330

Abstract

The World Health Organization (WHO) has set forth a global call for eradicating malaria, caused majorly by the protozoan parasites Plasmodium falciparum and Plasmodium vivax. The lack of diagnostic biomarkers for P. vivax, especially those that differentiate the parasite from P. falciparum, significantly hinders P. vivax elimination. Here, we show that P. vivax tryptophan-rich antigen (PvTRAg) can be a diagnostic biomarker for diagnosing P. vivax in malaria patients. We report that polyclonal antibodies against purified PvTRAg protein show interactions with purified PvTRAg and native PvTRAg using Western blots and indirect enzyme-linked immunosorbent assay (ELISA). We also developed an antibody-antigen-based qualitative assay using biolayer interferometry (BLI) to detect vivax infection using plasma samples from patients with different febrile diseases and healthy controls. The polyclonal anti-PvTRAg antibodies were used to capture free native PvTRAg from the patient plasma samples using BLI, providing a new expansion range to make the assay quick, accurate, sensitive, and high-throughput. The data presented in this report provides a proof of concept for PvTRAg, a new antigen, for developing a diagnostic assay for P. vivax identification and differentiation from the rest of the Plasmodium species and, at a later stage, translating the BLI assay into affordable, point-of-care formats to make it more accessible. Full article

(This article belongs to the Special Issue Novel Approaches for Diagnosis of Parasitic Disease)

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9 pages, 1135 KiB

Open AccessArticle

Accuracy Study of Kato-Katz and Helmintex Methods for Diagnosis of Schistosomiasis Mansoni in a Moderate Endemicity Area in Sergipe, Northeastern Brazil

by Daniel Lima Menezes, Carlos Thailan de Jesus Santos, Yvanna Louise Di Christine Oliveira, Vinícius Torres Castro Campos, Deborah Aparecida Negrão-Corrêa, Stefan Michael Geiger, José Rodrigo Santos Silva, Sona Jain, Luciana Maria Oliveira, Ricardo Toshio Fujiwara, Carlos Graeff-Teixeira and Silvio Santana Dolabella

Diagnostics 2023, 13(3), 527; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics13030527 - 31 Jan 2023

Cited by 5 | Viewed by 2000

Abstract

Schistosomiasis is a neglected tropical disease (NTD) caused by blood flukes from the genus Schistosoma. Brazil hosts the main endemic area in the Americas, where Schistosoma mansoni is the only species causing the disease. Kato-Katz (KK) thick smear is the WHO recommended screening test for populational studies, but there is growing evidence for the sensitivity limitations associated with KK, especially in areas with low parasite loads. Helmintex (HTX) is another highly sensitive egg-detection method, based on the magnetic properties of S. mansoni eggs and their isolation in a magnetic field. The objective of this study is to evaluate both KK and HTX in a moderate endemic locality, Areia Branca, located in the municipality of Pacatuba, in the state of Sergipe in northeastern Brazil. From 234 individual fecal samples, two KK thick smears were prepared and evaluated for each sample. Similarly, 30 g of each fecal sample was processed by HTX protocol. Eggs were detected in 80 (34.18%) residents. Twenty-three (9.83%) samples were positive for eggs (only by KK), and 77 (32.91%) samples showed positive for eggs (only by HTX). Sensitivity, specificity, and accuracy estimates gave values of 28.75%, 100% and 75.64%, respectively, for KK, and 96.25%, 100% and 98.72% respectively, for HTX. The positive predictive value was 100% for both methods, while the negative predictive value was 72.99% for KK and 98.09% for HTX. Overall, HTX presented a superior performance compared to the one sample, two slides KK examination. The study confirms the role of HTX as a reference method for the definition of true-positive samples in comparative accuracy studies and its potential role in the late stages when the certification of schistosomiasis transmission interruption is required. Diagnostic tests are important tools for the elimination of this NTD, besides the effective implementation of safe water, basic sanitation, snail control, and the treatment of infected populations. Full article

(This article belongs to the Special Issue Novel Approaches for Diagnosis of Parasitic Disease)

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13 pages, 2481 KiB

Open AccessArticle

The Humanised NPY-mRFP RBL Reporter Cell Line Is a Fast and Inexpensive Tool for Detection of Allergen-Specific IgE in Human Sera

by Prema S. Prakash, Nafal J. S. Barwary, Michael H. W. Weber, Daniel Wan, Iván Conejeros, Bernardo Pereira Moreira, Waleed S. Alharbi, Jaap J. van Hellemond, Jude Akinwale and Franco H. Falcone

Diagnostics 2022, 12(9), 2063; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12092063 - 25 Aug 2022

Viewed by 2093

Abstract

Rat basophilic leukaemia (RBL) cells have been used for decades as a model of high-affinity Immunoglobulin E (IgE) receptor (FcεRI) signalling. Here, we describe the generation and use of huNPY-mRFP, a new humanised fluorescent IgE reporter cell line. Fusion of Neuropeptide Y (NPY) with monomeric red fluorescent protein (mRFP) results in targeting of fluorescence to the granules and its fast release into the supernatant upon IgE-dependent stimulation. Following overnight sensitisation with serum, optimal release of fluorescence upon dose-dependent stimulation with allergen-containing extracts could be measured after 45 min, without cell lysis or addition of any reagents. Five substitutions (D194A, K212A, K216A, K226A, and K230A) were introduced into the FcεRIα cDNA used for transfection, which resulted in the removal of known endoplasmic reticulum retention signals and high surface expression of human FcεRIα* in huNPY-mRFP cells (where * denotes the penta-substituted variant), comparable to the ~500,000 FcεRIα molecules per cell in the RS-ATL8 humanised luciferase reporter, which is a human FcεRIα/FcεRIγ double transfectant. The huNPY-mRFP reporter was used to demonstrate engagement of specific IgE in sera of Echinococcus granulosus-infected individuals by E. granulosus elongation factor EgEF-1β and, to a lesser extent, by EgEF-1δ, which had been previously described as IgE-immunoreactive EgEF-1β/δ. Full article

(This article belongs to the Special Issue Novel Approaches for Diagnosis of Parasitic Disease)

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15 pages, 1857 KiB

Open AccessArticle

Colorimetric and Real-Time Loop-Mediated Isothermal Amplification (LAMP) for Detection of Loa loa DNA in Human Blood Samples

by Begoña Febrer-Sendra, Pedro Fernández-Soto, Beatriz Crego-Vicente, Juan García-Bernalt Diego, Thuy-Huong Ta-Tang, Pedro Berzosa, Rufino Nguema, Policarpo Ncogo, María Romay-Barja, Zaida Herrador, Agustín Benito and Antonio Muro

Diagnostics 2022, 12(5), 1079; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12051079 - 25 Apr 2022

Cited by 2 | Viewed by 2421

Abstract

Loiasis, caused by the filarial nematode Loa loa, is endemic in Central and West Africa. Loa loa has been associated with severe adverse reactions in high Loa-infected individuals receiving ivermectin during mass drug administration programs for the control of onchocerciasis and lymphatic filariasis. Diagnosis of loiasis still depends on microscopy in blood samples, but this is not effective for large-scale surveys. New diagnostics methods for loiasis are urgently needed. Previously, we developed a colorimetric high-sensitive and species-specific LAMP for Loa loa DNA detection. Here, we evaluate it in a set of 100 field-collected clinical samples stored as dried blood spots. In addition, Loa loa-LAMP was also evaluated in real-time testing and compared with microscopy and a specific PCR/nested PCR. A simple saponin/Chelex-based method was used to extract DNA. Colorimetric and real-time LAMP assays detected more samples with microscopy-confirmed Loa loa and Loa loa/Mansonella perstans mixed infections than PCR/nested-PCR. Samples with the highest Loa loa microfilariae counts were amplified faster in real-time LAMP assays. Our Loa loa-LAMP could be a promising molecular tool for the easy, rapid and accurate screening of patients for loiasis in endemic areas with low-resource settings. The real-time testing (feasible in a handheld device) could be very useful to rule out high-microfilariae loads in infected patients. Full article

(This article belongs to the Special Issue Novel Approaches for Diagnosis of Parasitic Disease)

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Review

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12 pages, 4573 KiB

Open AccessReview

Recent Advances and Potential Future Applications of MALDI-TOF Mass Spectrometry for Identification of Helminths

by Issa Sy, Lucie Conrad and Sören L. Becker

Diagnostics 2022, 12(12), 3035; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12123035 - 03 Dec 2022

Cited by 2 | Viewed by 1711

Abstract

Helminth infections caused by nematodes, trematodes, and cestodes are major neglected tropical diseases and of great medical and veterinary relevance. At present, diagnosis of helminthic diseases is mainly based on microscopic observation of different parasite stages, but microscopy is associated with limited diagnostic accuracy. Against this background, recent studies described matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry as a potential, innovative tool for helminth identification and differentiation. MALDI-TOF mass spectrometry is based on the analysis of spectra profiles generated from protein extracts of a given pathogen. It requires an available spectra database containing reference spectra, also called main spectra profiles (MSPs), which are generated from well characterized specimens. At present, however, there are no commercially available databases for helminth identification using this approach. In this narrative review, we summarize recent developments and published studies between January 2019 and September 2022 that report on the use of MALDI-TOF mass spectrometry for helminths. Current challenges and future research needs are identified and briefly discussed. Full article

(This article belongs to the Special Issue Novel Approaches for Diagnosis of Parasitic Disease)

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14 pages, 1226 KiB

Open AccessReview

Application of Proteomics to the Study of the Therapeutics and Pathogenicity of Giardia duodenalis

by Ahmad Fudail Eiyad Aziz, Norhamizah Roshidi, Nurulhasanah Othman, Khayriyyah Mohd Hanafiah and Norsyahida Arifin

Diagnostics 2022, 12(11), 2744; https://0-doi-org.brum.beds.ac.uk/10.3390/diagnostics12112744 - 09 Nov 2022

Cited by 2 | Viewed by 1559

Abstract

Giardia duodenalis remains a neglected tropical disease. A key feature of the sustained transmission of Giardia is the ability to form environmentally resistant cysts. For the last 38 years, proteomics has been utilised to study various aspects of the parasite across different life cycle stages. Thirty-one articles have been published in PubMed from 2012 to 2022 related to the proteomics of G. duodenalis. Currently, mass spectrometry with LC-MS/MS and MALDI-TOF/TOF has been commonly utilised in proteomic analyses of Giardia, which enables researchers to determine potential candidates for diagnostic biomarkers as well as vaccine and drug targets, in addition to allowing them to investigate the virulence of giardiasis, the pathogenicity mechanisms of G. duodenalis, and the post-translational modifications of Giardia proteins throughout encystation. Over the last decade, valuable information from proteomics analyses of G. duodenalis has been discovered in terms of the pathogenesis and virulence of Giardia, which may provide guidance for the development of better means with which to prevent and reduce the impacts of giardiasis. Nonetheless, there is room for improving proteomics analyses of G. duodenalis, since genomic sequences for additional assemblages of Giardia have uncovered previously unknown proteins associated with the Giardia proteome. Therefore, this paper aims to review the applications of proteomics for the characterisation of G. duodenalis pathogenicity and the discovery of novel vaccine as well as drug targets, in addition to proposing some general directions for future Giardia proteomic research. Full article

(This article belongs to the Special Issue Novel Approaches for Diagnosis of Parasitic Disease)

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