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1.
Virus Genes ; 2022 May 10.
Article in English | MEDLINE | ID: mdl-35536436

ABSTRACT

Dengue is a rapidly spreading arboviral disease that can be transmitted through any of the four types of dengue virus (DENV) serotypes. Previous studies have observed that individuals who have a pre-existing secondary infection due to a different dengue serotype, experience severe forms of this disease. During a DENV outbreak, a time-sensitive preliminary diagnosis of the origin of DENV might be useful in controlling the epidemic. Here, we developed a rapid and accurate one-step TB Green RT-PCR-based high-resolution melting (HRM) assay to identify and serotype DENV using serotyping primers based on the alignment with the E gene. This assay had a detection limit of 7.7 × 102 (DENV 1), 3.8 × 102 (DENV 2), 6.2 × 102 (DENV 3), and 1.2 × 103 (DENV 4) RNA copies/mL. No cross-reactivity with the Chikungunya, Zika, and Japanese encephalitis viruses was observed. The feasibility of using this assay for clinical diagnosis was evaluated in DENV-positive patient sera. The HRM assay and the RT-qPCR had complete matched results derived from DENV detection, including 51 serum positive and 20 serum negative. Additionally, eight DENV 2 strains in the same serotype were successfully differentiated by an HRM assay. Thus, this assay facilitated accurate detection and serotyping of DENV, along with the time-sensitive identification of the infectious focus of different DENVs.

2.
J Perinat Med ; 2022 Apr 18.
Article in English | MEDLINE | ID: mdl-35427445

ABSTRACT

OBJECTIVES: To identify the prevalence of viral congenital infections in newborns classified as premature, low-birthweight, small for gestational age or intrauterine growth restriction. METHODS: The definition considered for selecting papers were: P as newborns younger than 28 days; V as low-birthweight, prematurity and intrauterine growth restriction; O as frequency of congenital infections with Cytomegalovirus, Parvovirus B19, Herpes Simplex, and Zika virus. The research was performed using EMBASE, LILACS, SCOPUS and MEDLINE databases, with no limitations on date and language. RESULTS: Eight studies were included. Manuscripts including Herpes Simplex, Zika virus or Parvovirus B19 did not fulfill the defined criteria. A wide variation in the frequency of CMV congenital infection (0-4.8%) was found, which might be attributed to regional and methodological differences between investigations. CONCLUSIONS: Newborn characteristics associated with CMV congenital infections may direct investigations towards these patients with a higher probability of infection. However, as data are controversial, studies concerning screening of infection are important to define recommendations of diagnosis.

3.
PLoS One ; 17(4): e0265820, 2022.
Article in English | MEDLINE | ID: mdl-35395015

ABSTRACT

INTRODUCTION: The rapid expansion of the novel SARS-CoV-2 virus has raised serious public health concerns due to the possibility of misdiagnosis in regions where arboviral diseases are endemic. We performed the first study in northern Peru to describe the detection of SARS-CoV-2 IgM antibodies in febrile patients with a suspected diagnosis of dengue and chikungunya fever. MATERIALS AND METHODS: A consecutive cross-sectional study was performed in febrile patients attending primary healthcare centers from April 2020 through March 2021. Patients enrolled underwent serum sample collection for the molecular and serological detection of DENV and CHIKV. Also, serological detection of IgM antibodies against SARS-CoV-2 was performed. RESULTS: 464 patients were included during the study period, of which (40.51%) were positive for one pathogen, meanwhile (6.90%) presented co-infections between 2 or more pathogens. The majority of patients with monoinfections were positive for SARS-CoV-2 IgM with (73.40%), followed by DENV 18.09% and CHIKV (8.51%). The most frequent co-infection was DENV + SARS-CoV-2 with (65.63%), followed by DENV + CHIKV and DENV + CHIKV + SARS-CoV-2, both with (12.50%). The presence of polyarthralgias in hands (43.75%, p<0.01) and feet (31.25%, p = 0.05) were more frequently reported in patients with CHIKV monoinfection. Also, conjunctivitis was more common in patients positive for SARS-CoV-2 IgM (11.45%, p<0.01). The rest of the symptoms were similar among all the study groups. CONCLUSION: SARS-CoV-2 IgM antibodies were frequently detected in acute sera from febrile patients with a clinical suspicion of arboviral disease. The presence of polyarthralgias in hands and feet may be suggestive of CHIKV infection. These results reaffirm the need to consider SARS-CoV-2 infection as a main differential diagnosis of acute febrile illness in arboviruses endemic areas, as well as to consider co-infections between these pathogens.


Subject(s)
COVID-19 , Chikungunya Fever , Chikungunya virus , Coinfection , Dengue Virus , Dengue , Zika Virus Infection , Antibodies, Viral , Arthralgia , COVID-19/diagnosis , COVID-19/epidemiology , Chikungunya Fever/diagnosis , Chikungunya Fever/epidemiology , Coinfection/diagnosis , Coinfection/epidemiology , Cross-Sectional Studies , Dengue/diagnosis , Dengue/epidemiology , Fever/diagnosis , Humans , Immunoglobulin M , Peru/epidemiology , SARS-CoV-2 , Zika Virus Infection/epidemiology
4.
Biosens Bioelectron ; 210: 114302, 2022 Apr 25.
Article in English | MEDLINE | ID: mdl-35487137

ABSTRACT

The recent worldwide spread of viral infections has highlighted the need for accurate, fast, and inexpensive disease diagnosis and monitorization methods. Current diagnostics tend to focus either on molecular or serological testing. In this work we propose a dual detection assay approach for viral diseases, where both serological and molecular assays are combined in a single analysis performed on a magnetoresistive system. This type of assay guarantees an accurate assessment of the infection phase, saving time and costs associated with multiple independent tests. Zika and dengue viruses were used as model diseases for the validation of the system. Human IgG anti-zika and anti-dengue antibodies were successfully detected in infected patients' serum, using a novel approach combining competitive and sandwich strategies in a magnetoresistive portable platform. Specificity and sensitivity values of 100% were obtained. Calibration curves with dynamic ranges between 10 ng/mL and 1 µg/mL were established achieving LODs of 1.26 and 1.38 nM for IgG anti-ZIKV and anti-DENV antibodies, respectively. Viral RNA detection down to a few hundreds of pM was also successfully carried out after the design of specific oligo probes and primers for RT-PCR amplification. Dual assays were performed for both viruses, where viral RNA and anti-virus antibodies in serum samples were simultaneously detected. The results obtained for the detection of the molecular and serological targets in the dual assay format show no significant difference between the ones obtained individually, proving the feasibility and accuracy of the dual detection assay. This assay format represents a new paradigm in viral infections diagnostics.

5.
J Clin Med ; 11(5)2022 Mar 01.
Article in English | MEDLINE | ID: mdl-35268441

ABSTRACT

Zika virus (ZIKV) was discovered in Uganda in 1947 and was originally isolated only in Africa and Asia. After a spike of microcephaly cases in Brazil, research has closely focused on different aspects of congenital ZIKV infection. In this review, we evaluated many aspects of the disease in order to build its natural history, with a focus on the long-term clinical and neuro-radiological outcomes in children. The authors have conducted a wide-ranging search spanning the 2012-2021 period from databases PubMed, PubMed Central, Web of Science, Medline, Scopus. Different sections reflect different points of congenital ZIKV infection syndrome: pathogenesis, prenatal diagnosis, clinical signs, neuroimaging and long-term developmental outcomes. It emerged that pathogenesis has not been fully clarified and that the clinical signs are not only limited to microcephaly. Given the current absence of treatments, we proposed schemes to optimize diagnostic protocols in endemic countries. It is essential to know the key aspects of this disease to guarantee early diagnosis, even in less severe cases, and an adequate management of the main chronic problems. Considering the relatively recent discovery of this congenital infectious syndrome, further studies and updated long-term follow-up are needed to further improve management strategies for this disease.

6.
ACS Biomater Sci Eng ; 8(3): 964-1000, 2022 03 14.
Article in English | MEDLINE | ID: mdl-35229605

ABSTRACT

Graphene- and carbon-based nanomaterials are key materials to develop advanced biosensors for the sensitive detection of many biomarkers owing to their unique properties. Biosensors have attracted increasing interest because they allow efficacious, sensitive, selective, rapid, and low-cost diagnosis. Biosensors are analytical devices based on receptors for the process of detection and transducers for response measuring. Biosensors can be based on electrochemical, piezoelectric, thermal, and optical transduction mechanisms. Early virus identification provides critical information about potentially effective and selective therapies, extends the therapeutic window, and thereby reduces morbidity. The sensitivity and selectivity of graphene can be amended via functionalizing it or conjoining it with further materials. Amendment of the optical and electrical features of the hybrid structure by introducing appropriate functional groups or counterparts is especially appealing for quick and easy-to-use virus detection. Various techniques for the electrochemical detection of viruses depending on antigen-antibody interactions or DNA hybridization are discussed in this work, and the reasons behind using graphene and related carbon nanomaterials for the fabrication are presented and discussed. We review the existing state-of-the-art directions of graphene-based classifications for detecting DNA, protein, and hormone biomarkers and summarize the use of the different biosensors to detect several diseases, like cancer, Alzheimer's disease, and diabetes, to sense numerous viruses, including SARS-CoV-2, human immunodeficiency virus, rotavirus, Zika virus, and hepatitis B virus, and to detect the recent pandemic virus COVID-19. The general concepts, mechanisms of action, benefits, and disadvantages of advanced virus biosensors are discussed to afford beneficial evidence of the creation and manufacture of innovative virus biosensors. We emphasize that graphene-based nanomaterials are ideal candidates for electrochemical biosensor engineering due to their special and tunable physicochemical properties.


Subject(s)
Biosensing Techniques , COVID-19 , Graphite , Nanostructures , Zika Virus Infection , Zika Virus , Biosensing Techniques/methods , Carbon , Electrochemical Techniques/methods , Humans , Nanostructures/chemistry , SARS-CoV-2
7.
Washington, D.C.; PAHO; 2022-03-24.
in English | PAHO-IRIS | ID: phr-55867

ABSTRACT

Evidence-based guidelines are one of the most useful tools for improving public health and clinical practice. Their purpose is to formulate interventions based on strong evidence of efficacy, avoid unnecessary risks, use resources efficiently, reduce clinical variability and, in essence, improve health and ensure quality care, which is the purpose of health systems and services. These guidelines were developed following the GRADE methodology, with the support of a panel of clinical experts from different countries, all convened by the Pan American Health Organization. By responding to twelve key questions about the clinical diagnosis and treatment of dengue, chikungunya, and Zika, evidence-based recommendations were formulated for pediatric, youth, adult, older adult, and pregnant patients who are exposed to these diseases or have a suspected or confirmed diagnosis of infection. The purpose of the guidelines is to prevent progression to severe forms of these diseases and the fatal events they may cause. The recommendations are intended for health professionals, including general, resident, and specialist physicians, nursing professionals, and medical and nursing students, who participate in caring for patients with suspected dengue, chikungunya, or Zika. They are also intended for health unit managers and the executive teams of national arboviral disease prevention and control programs, who are responsible for facilitating the process of implementing these guidelines.


Subject(s)
Chikungunya virus , Dengue Virus , Communicable Diseases , Clinical Diagnosis , Dengue , Zika Virus , Arboviruses , Pregnant Women , Child , Pediatrics , Aged , Communicable Disease Control
8.
Lab Chip ; 22(9): 1748-1763, 2022 May 03.
Article in English | MEDLINE | ID: mdl-35357372

ABSTRACT

This paper introduces a digital microfluidic (DMF) platform for portable, automated, and integrated Zika viral RNA extraction and amplification. The platform features reconfigurable DMF cartridges offering a closed, humidified environment for sample processing at elevated temperatures, as well as programmable control instrumentation with a novel thermal cycling unit regulated using a proportional integral derivative (PID) feedback loop. The system operates on 12 V DC power, which can be supplied by rechargeable battery packs for remote testing. The DMF system was optimized for an RNA processing pipeline consisting of the following steps: 1) magnetic-bead based RNA extraction from lysed plasma samples, 2) RNA clean-up, and 3) integrated, isothermal amplification of Zika RNA. The DMF pipeline was coupled to a paper-based, colorimetric cell-free protein expression assay for amplified Zika RNA mediated by toehold switch-based sensors. Blinded laboratory evaluation of Zika RNA spiked in human plasma yielded a sensitivity and specificity of 100% and 75% respectively. The platform was then transported to Recife, Brazil for evaluation with infectious Zika viruses, which were detected at the 100 PFU mL-1 level from a 5 µL sample (equivalent to an RT-qPCR cycle threshold value of 32.0), demonstrating its potential as a sample processing platform for miniaturized diagnostic testing.


Subject(s)
Zika Virus Infection , Zika Virus , Humans , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , RNA , RNA, Viral/analysis , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Specimen Handling , Zika Virus/genetics , Zika Virus Infection/diagnosis
9.
Nat Biomed Eng ; 6(3): 246-256, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35256758

ABSTRACT

In low-resource settings, resilience to infectious disease outbreaks can be hindered by limited access to diagnostic tests. Here we report the results of double-blinded studies of the performance of paper-based diagnostic tests for the Zika and chikungunya viruses in a field setting in Latin America. The tests involved a cell-free expression system relying on isothermal amplification and toehold-switch reactions, a purpose-built portable reader and onboard software for computer vision-enabled image analysis. In patients suspected of infection, the accuracies and sensitivities of the tests for the Zika and chikungunya viruses were, respectively, 98.5% (95% confidence interval, 96.2-99.6%, 268 serum samples) and 98.5% (95% confidence interval, 91.7-100%, 65 serum samples) and approximately 2 aM and 5 fM (both concentrations are within clinically relevant ranges). The analytical specificities and sensitivities of the tests for cultured samples of the viruses were equivalent to those of the real-time quantitative PCR. Cell-free synthetic biology tools and companion hardware can provide de-centralized, high-capacity and low-cost diagnostics for use in low-resource settings.


Subject(s)
Chikungunya Fever , Chikungunya virus , Dengue , Zika Virus Infection , Zika Virus , Chikungunya Fever/diagnosis , Chikungunya Fever/epidemiology , Dengue/diagnosis , Humans , Zika Virus/genetics , Zika Virus Infection/diagnosis , Zika Virus Infection/epidemiology
10.
J Med Virol ; 94(6): 2528-2536, 2022 06.
Article in English | MEDLINE | ID: mdl-35146775

ABSTRACT

Due to the concurrent prevalence and increasing risk of coinfection of the clinically important Arboviruses, timely and accurate differential diagnosis is important for clinical management and the epidemiological investigation. A two-tube multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR) assay for the simultaneous detection of Zika virus (ZIKV), chikungunya virus (CHIKV), dengue virus (DENV), yellow fever virus (YFV), West Nile virus (WNV), and Japanese encephalitis virus (JEV) was developed and optimized with high specificity and sensitivity. The detection limit for all the six viruses could reach as low as five genome equivalent copies and 2.8 × 10- 3 tissue culture infectious doses (TCID50 ) for ZIKV, YFV, CHIKV and 2.8 × 10-2 TCID50  for JEV per reaction, with high accuracy and precision (R2 > 0.99). The coefficient of variation of intra-assay and inter-assay for our quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was low, and the obtained positive rates ad Ct values of this assay were comparable with singleplex commercial kits. Moreover, the multiplex qRT-PCR assay was able to detect possible co-infections without competitive inhibition of target viral genomes. In conclusion, our rapid, sensitive, cost-effective multiplex qRT-PCR will be of great use for differential diagnosis in a clinical setting and epidemiological investigation during surveillance.


Subject(s)
Chikungunya Fever , Chikungunya virus , Dengue Virus , Dengue , Encephalitis Virus, Japanese , Encephalitis Viruses, Japanese , West Nile Fever , Yellow Fever , Zika Virus Infection , Zika Virus , Chikungunya Fever/diagnosis , Chikungunya virus/genetics , Dengue/diagnosis , Dengue Virus/genetics , Encephalitis Virus, Japanese/genetics , Encephalitis Viruses, Japanese/genetics , Humans , Reverse Transcriptase Polymerase Chain Reaction , West Nile Fever/diagnosis , Yellow Fever/diagnosis , Yellow fever virus/genetics , Zika Virus/genetics
11.
Viruses ; 14(2)2022 02 08.
Article in English | MEDLINE | ID: mdl-35215941

ABSTRACT

Flavivirus outbreaks require fast and reliable diagnostics that can be easily adapted to newly emerging and re-emerging flaviviruses. Due to the serological cross-reactivity among flavivirus antibodies, neutralization tests (NT) are considered the gold standard for sero-diagnostics. Here, we first established wild-type single-round infectious virus replicon particles (VRPs) by packaging a yellow fever virus (YFV) replicon expressing Gaussia luciferase (Gluc) with YFV structural proteins in trans using a double subgenomic Sindbis virus (SINV) replicon. The latter expressed the YFV envelope proteins prME via the first SINV subgenomic promoter and the capsid protein via a second subgenomic SINV promoter. VRPs were produced upon co-electroporation of replicon and packaging RNA. Introduction of single restriction enzyme sites in the packaging construct flanking the prME sequence easily allowed to exchange the prME moiety resulting in chimeric VRPs that have the surface proteins of other flaviviruses including dengue virus 1--4, Zika virus, West Nile virus, and tick-borne encephalitis virus. Besides comparing the YF-VRP based NT assay to a YF reporter virus NT assay, we analyzed the neutralization efficiencies of different human anti-flavivirus sera or a monoclonal antibody against all established VRPs. The assays were performed in a 96-well high-throughput format setting with Gluc as readout in comparison to classical plaque reduction NTs indicating that the VRP-based NT assays are suitable for high-throughput analyses of neutralizing flavivirus antibodies.


Subject(s)
Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Flavivirus/immunology , High-Throughput Screening Assays/methods , Cross Reactions , Flavivirus/classification , Flavivirus/genetics , Flavivirus/physiology , Genes, Reporter , Luciferases/genetics , Luciferases/metabolism , Neutralization Tests , Replicon , Sindbis Virus/genetics , Sindbis Virus/immunology , Sindbis Virus/physiology , Virion/genetics , Virion/immunology , Virion/physiology , Yellow fever virus/genetics , Yellow fever virus/immunology , Yellow fever virus/physiology
12.
PLoS One ; 17(2): e0262149, 2022.
Article in English | MEDLINE | ID: mdl-35139078

ABSTRACT

There is an urgent need for better diagnostic and analytical methods for vaccine research and infection control in virology. This has been highlighted by recently emerging viral epidemics and pandemics (Zika, SARS-CoV-2), and recurring viral outbreaks like the yellow fever outbreaks in Angola and the Democratic Republic of Congo (2016) and in Brazil (2016-2018). Current assays to determine neutralising activity against viral infections in sera are costly in time and equipment and suffer from high variability. Therefore, both basic infection research and diagnostic population screenings would benefit from improved methods to determine virus-neutralising activity in patient samples. Here we describe a robust, objective, and scalable Fluorescence Reduction Neutralisation Test (FluoRNT) for yellow fever virus, relying on flow cytometric detection of cells infected with a fluorescent Venus reporter containing variant of the yellow fever vaccine strain 17D (YF-17D-Venus). It accurately measures neutralising antibody titres in human serum samples within as little as 24 h. Samples from 32 vaccinees immunised with YF-17D were tested for neutralising activity by both a conventional focus reduction neutralisation test (FRNT) and FluoRNT. Both types of tests proved to be equally reliable for the detection of neutralising activity, however, FluoRNT is significantly more precise and reproducible with a greater dynamic range than conventional FRNT. The FluoRNT assay protocol is substantially faster, easier to control, and cheaper in per-assay costs. FluoRNT additionally reduces handling time minimising exposure of personnel to patient samples. FluoRNT thus brings a range of desirable features that can accelerate and standardise the measurement of neutralising anti-yellow fever virus antibodies. It could be used in applications ranging from vaccine testing to large cohort studies in systems virology and vaccinology. We also anticipate the potential to translate the methodology and analysis of FluoRNT to other flaviviruses such as West Nile, Dengue and Zika or to RNA viruses more generally.


Subject(s)
Antibodies, Neutralizing/immunology , Yellow Fever/immunology , Yellow fever virus/immunology , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Antibodies, Viral/immunology , Chlorocebus aethiops , Fluorescence , Humans , Neutralization Tests/economics , Neutralization Tests/methods , Vero Cells , Yellow Fever/blood , Yellow Fever/virology
13.
PLoS Negl Trop Dis ; 16(2): e0010152, 2022 02.
Article in English | MEDLINE | ID: mdl-35120141

ABSTRACT

BACKGROUND: Chikungunya virus (CHIKV) causes febrile illnesses and has always been misdiagnosed as other viral infections, such as dengue and Zika; thus, a laboratory test is needed. Serological tests are commonly used to diagnose CHIKV infection, but their accuracy is questionable due to varying degrees of reported sensitivities and specificities. Herein, we conducted a systematic review and meta-analysis to evaluate the diagnostic accuracy of serological tests currently available for CHIKV. METHODOLOGY AND PRINCIPAL FINDINGS: A literature search was performed in PubMed, CINAHL Complete, and Scopus databases from the 1st December 2020 until 22nd April 2021. Studies reporting sensitivity and specificity of serological tests against CHIKV that used whole blood, serum, or plasma were included. QUADAS-2 tool was used to assess the risk of bias and applicability, while R software was used for statistical analyses. Thirty-five studies were included in this meta-analysis; 72 index test data were extracted and analysed. Rapid and ELISA-based antigen tests had a pooled sensitivity of 85.8% and 82.2%, respectively, and a pooled specificity of 96.1% and 96.0%, respectively. According to our meta-analysis, antigen detection tests serve as a good diagnostic test for acute-phase samples. The IgM detection tests had more than 90% diagnostic accuracy for ELISA-based tests, immunofluorescence assays, in-house developed tests, and samples collected after seven days of symptom onset. Conversely, low sensitivity was found for the IgM rapid test (42.3%), commercial test (78.6%), and for samples collected less than seven of symptom onset (26.2%). Although IgM antibodies start to develop on day 2 of CHIKV infection, our meta-analysis revealed that the IgM detection test is not recommended for acute-phase samples. The diagnostic performance of the IgG detection tests was more than 93% regardless of the test formats and whether the test was commercially available or developed in-house. The use of samples collected after seven days of symptom onset for the IgG detection test suggests that IgG antibodies can be detected in the convalescent-phase samples. Additionally, we evaluated commercial IgM and IgG tests for CHIKV and found that ELISA-based and IFA commercial tests manufactured by Euroimmun (Lübeck, Germany), Abcam (Cambridge, UK), and Inbios (Seattle, WA) had diagnostic accuracy of above 90%, which was similar to the manufacturers' claim. CONCLUSION: Based on our meta-analysis, antigen or antibody-based serological tests can be used to diagnose CHIKV reliably, depending on the time of sample collection. The antigen detection tests serve as a good diagnostic test for samples collected during the acute phase (≤7 days post symptom onset) of CHIKV infection. Likewise, IgM and IgG detection tests can be used for samples collected in the convalescent phase (>7 days post symptom onset). In correlation to the clinical presentation of the patients, the combination of the IgM and IgG tests can differentiate recent and past infections.


Subject(s)
Antigens, Viral/isolation & purification , Chikungunya Fever/diagnosis , Serologic Tests/standards , Antigens, Viral/blood , Chikungunya virus/immunology , Chikungunya virus/isolation & purification , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Sensitivity and Specificity
14.
Front Immunol ; 13: 793882, 2022.
Article in English | MEDLINE | ID: mdl-35154111

ABSTRACT

In a world with an increasing population at risk of exposure to arthropod-borne flaviviruses, access to timely and accurate diagnostic tests would impact profoundly on the management of cases. Twenty peptides previously identified using a flavivirus proteome-wide microarray were evaluated to determine their discriminatory potential to detect dengue virus (DENV) infection. This included nine peptides recognized by IgM antibodies (PM peptides) and 11 peptides recognized by IgG antibodies (PG peptides). A bead-based multiplex peptide immunoassay (MPIA) using the Luminex technology was set-up to determine Ab binding levels to each of these peptides in a panel of 323 carefully selected human serum samples. Sera are derived from individuals either infected with different viruses, namely, the four DENV serotypes, Zika virus (ZIKV), yellow fever virus (YFV), chikungunya virus (CHIKV), West Nile virus (WNV) and Human immunodeficiency virus (HIV), or receiving vaccination against YFV, tick-borne encephalitis (TBEV), and Japanese encephalitis virus (JEV). Additionally, a set of healthy controls were included. We targeted a minimum specificity of 80% for all the analysis. The PG-9 peptide had the best sensitivity (73%) when testing DENV sera from acute patients (A-DENV; <8 days since symptom onset). With sera from convalescent DENV patients (C-DENV; >10 days since symptom onset) the FPG-1 peptide was the best seromarker with a sensitivity of 86%. When combining all A-DENV and C-DENV samples, peptides PM-22 and FPG-1 had the best-diagnostic performance with a sensitivity of 60 and 61.1%, and areas under the curve (AUC) of 0.7865 and 0.8131, respectively. A Random forest (RF) algorithm was used to select the best combination of peptides to classify DENV infection at a targeted specificity >80%. The best RF model for PM peptides that included A-DENV and C-DENV samples, reached a sensitivity of 72.3%, while for PG peptides, the best RF models for A-DENV only, C-DENV only and A-DENV + C-DENV reached a sensitivity of 88.9%, 89.1%, and 88.3%, respectively. In conclusion, the combination of multiple peptides constitutes a founding set of seromarkers for the discrimination of DENV infected individuals from other flavivirus infections.


Subject(s)
Biomarkers , Dengue Virus/physiology , Dengue/diagnosis , Dengue/microbiology , Peptides , Viral Proteins , Adolescent , Adult , Aged , Antibodies, Viral , Biomarkers/blood , Child , Child, Preschool , Dengue/blood , Dengue/epidemiology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Male , Middle Aged , Peptides/blood , Peru/epidemiology , Prognosis , Proteome , Proteomics/methods , ROC Curve , Reagent Kits, Diagnostic , Reproducibility of Results , Viral Proteins/blood , Young Adult
15.
Braz J Microbiol ; 2022 Feb 07.
Article in English | MEDLINE | ID: mdl-35129818

ABSTRACT

Dengue is the most prevalent arboviral disease in humans in tropical and subtropical regions, especially in urban areas, and can cause major epidemics. Although a self-limiting illness, it may sometimes have serious hemorrhagic manifestations, and the outcome of dengue hemorrhagic fever has similar clinical manifestations as in other infections, which could result in death. Therefore, autopsy procedures are required under certain circumstances such as in hemorrhagic fevers, sometimes to confirm or to clarify the diagnosis that may have epidemiological consequences. Normally, the Immunohistochemistry Laboratory of the Pathology Center of Adolfo Lutz Institute receives autopsy samples from different hospitals in Sao Paulo State to confirm a previous diagnosis, especially hemorrhagic fever of infectious etiology. For this diagnosis, we have been using a mouse polyclonal antibody to dengue virus that often does not provide a clear conclusion, because of background staining or no relevant immunostaining, which hampers the histopathological analysis. Accordingly, in the present study, anti-DENV-NS1 monoclonal antibody (4H2) was tested to determine its accuracy in immunohistochemical analysis. Twenty-four autopsy cases of hemorrhagic febrile syndrome showing histopathological alterations compatible with dengue disease were studied: twenty cases were confirmed by RT-PCR for DENV-2 and in four by RT-PCR for yellow fever virus. Samples from autopsied cases of deaths caused by other infectious diseases (two meningitis C and two severe acute respiratory syndrome caused by influenza A H1N1) were included as negative control cases. Positive immunostaining for DENV-NS1 was detected in 16/20 (80%) liver samples and 11/15 (73%) spleen samples from autopsied hemorrhagic dengue patients, whereas the polyclonal antibody detected DENV antigens in 12/20 (60%) liver and in 6/15 (40%) spleen samples from the same cases. Positive results were not obtained with liver biopsy samples from yellow fever or Neisseria meningitides and Flu-A cases. 4H2 mAb recognizes the native protein of the four DENV serotypes in infected cells and did not cross-react with native ZIKV- or CHKV-infected cells by immunohistochemical assay, so it is a useful tool for differential histopathological conclusion of acute febrile hemorrhagic deaths.

17.
Int J Infect Dis ; 119: 34-37, 2022 Jun.
Article in English | MEDLINE | ID: mdl-34990800

ABSTRACT

INTRODUCTION: Yellow fever (YF) is a public health threat with frequent outbreaks in tropical and subtropical areas, despite the existence of a safe and effective vaccine. The diagnosis of acute infection of the etiologic agent relies mainly on real-time reverse transcription-polymerase chain reaction (RT-qPCR)-based assays. OBJECTIVES: The aim of this study was to evaluate and compare this novel protocol for yellow fever virus (YFV) diagnosis against assays developed in-house by reference laboratories for arboviruses. METHODS: We developed a novel molecular protocol for the detection of YFV that includes an Internal Control to validate the reaction and an External Control to monitor the RNA extraction efficiency. RESULTS AND DISCUSSION: Our assay detects one viral genome per reaction and displays no cross-reactions with dengue (1-4), Zika, or Chikungunya viruses. This novel assay yielded 95% of agreement with the reference method recommended by the Pan American Health Organization when analyzing 204 clinical samples and cultured viruses, these samples were analyzed in 3 different diagnosis centers for arboviruses in Brazil. The data suggest the use of the proposed multiplex assay protocol to do routine tests in a clinical laboratory. This product adds higher specificity and sensitivity in addition to reduced cost per test due to hands-on time and reagent spending.

18.
Washington D.C; Organización Panamericana de la Salud; 1 ed; Ene. 2022. 16 p. ilus.
Monography in Spanish | MINSAPERÚ, LIPECS, MINSAPERÚ | ID: biblio-1352698

ABSTRACT

La presente publicación describe las intervenciones formuladas con sólidas pruebas de eficacia, evitar riesgos innecesarios, utilizar los recursos de forma eficiente, disminuir la variabilidad clínica y, en esencia, mejorar la salud y garantizar una atención de calidad, razón de ser de los sistemas y servicios de salud. Las presentes directrices se elaboraron siguiendo la metodología GRADE con el apoyo de un panel de expertos clínicos de distintos países, todos ellos convocados por la Organización Panamericana de la Salud. Por medio de la respuesta a doce preguntas clave sobre el diagnóstico clínico y el tratamiento del dengue, el chikunguña y el zika, se formulan recomendaciones basadas en evidencia para pacientes pediátricos, jóvenes, adultos, personas mayores y embarazadas expuestos a estas enfermedades o con sospecha o diagnóstico confirmado de infección.


Subject(s)
Quality of Health Care , Health Surveillance , Clinical Diagnosis , Epidemiology , Dengue , Chikungunya Fever
19.
Washington, D.C.; OPS; 2022-01-03.
in Spanish | PAHO-IRIS | ID: phr-55125

ABSTRACT

En la actualidad, las guías basadas en la evidencia constituyen una de las herramientas más útiles para mejorar la salud pública y la práctica clínica. Su finalidad es formular intervenciones con sólidas pruebas de eficacia, evitar riesgos innecesarios, utilizar los recursos de forma eficiente, disminuir la variabilidad clínica y, en esencia, mejorar la salud y garantizar una atención de calidad, razón de ser de los sistemas y servicios de salud. Las presentes directrices se elaboraron siguiendo la metodología GRADE con el apoyo de un panel de expertos clínicos de distintos países, todos ellos convocados por la Organización Panamericana de la Salud. Por medio de la respuesta a doce preguntas clave sobre el diagnóstico clínico y el tratamiento del dengue, el chikunguña y el zika, se formulan recomendaciones basadas en evidencia para pacientes pediátricos, jóvenes, adultos, personas mayores y embarazadas expuestos a estas enfermedades o con sospecha o diagnóstico confirmado de infección. La finalidad de las directrices es evitar la progresión a las formas graves y a los eventos mortales que puedan causar. Las recomendaciones están dirigidas a profesionales de la salud, incluidos el personal médico general, residente y especialista; y los profesionales de enfermería, así como a estudiantes de medicina y enfermería, quienes de una u otra forma participan en la atención de pacientes con sospecha de dengue, chikunguña o zika. También se dirige a los administradores de las unidades de salud y a los equipos directivos de los programas nacionales de prevención y control de enfermedades arbovirales, quienes tienen la responsabilidad de facilitar el proceso de aplicación de estas directrices. Esperamos que esta publicación beneficie no solo al personal de salud, que dispondrá de información científica actualizada y de la mejor calidad posible, sino a los menores, los adultos, las embarazadas, las personas mayores y la población en general, quienes recibirán una mejor atención de salud prestada por personal médico debidamente capacitado.


Subject(s)
Chikungunya virus , Dengue Virus , Communicable Diseases , Clinical Diagnosis , Dengue , Zika Virus , Arboviruses , Pregnant Women , Child , Pediatrics , Aged , Communicable Disease Control
20.
Washington; Organización Panamericana de la Salud; rev; Jan. 2022, 03. 158 p. tab.
Monography in English, Spanish | LILACS, BIGG - GRADE guidelines | ID: biblio-1362814

ABSTRACT

En la actualidad, las guías basadas en la evidencia constituyen una de las herramientas más útiles para mejorar la salud pública y la práctica clínica. Su finalidad es formular intervenciones con sólidas pruebas de eficacia, evitar riesgos innecesarios, utilizar los recursos de forma eficiente, disminuir la variabilidad clínica y, en esencia, mejorar la salud y garantizar una atención de calidad, razón de ser de los sistemas y servicios de salud. Las presentes directrices se elaboraron siguiendo la metodología GRADE con el apoyo de un panel de expertos clínicos de distintos países, todos ellos convocados por la Organización Panamericana de la Salud. Por medio de la respuesta a doce preguntas clave sobre el diagnóstico clínico y el tratamiento del dengue, el chikunguña y el zika, se formulan recomendaciones basadas en evidencia para pacientes pediátricos, jóvenes, adultos, personas mayores y embarazadas expuestos a estas enfermedades o con sospecha o diagnóstico confirmado de infección. La finalidad de las directrices es evitar la progresión a las formas graves y a los eventos mortales que puedan causar. Las recomendaciones están dirigidas a profesionales de la salud, incluidos el personal médico general, residente y especialista; y los profesionales de enfermería, así como a estudiantes de medicina y enfermería, quienes de una u otra forma participan en la atención de pacientes con sospecha de dengue, chikunguña o zika. También se dirige a los administradores de las unidades de salud y a los equipos directivos de los programas nacionales de prevención y control de enfermedades arbovirales, quienes tienen la responsabilidad de facilitar el proceso de aplicación de estas directrices. Esperamos que esta publicación beneficie no solo al personal de salud, que dispondrá de información científica actualizada y de la mejor calidad posible, sino a los menores, los adultos, las embarazadas, las personas mayores y la población en general, quienes recibirán una mejor atención de salud prestada por personal médico debidamente capacitado.


Evidence-based guidelines are one of the most useful tools for improving public health and clinical practice. Their purpose is to formulate interventions based on strong evidence of efficacy, avoid unnecessary risks, use resources efficiently, reduce clinical variability and, in essence, improve health and ensure quality care, which is the purpose of health systems and services. These guidelines were developed following the GRADE methodology, with the support of a panel of clinical experts from different countries, all convened by the Pan American Health Organization. By responding to twelve key questions about the clinical diagnosis and treatment of dengue, chikungunya, and Zika, evidence-based recommendations were formulated for pediatric, youth, adult, older adult, and pregnant patients who are exposed to these diseases or have a suspected or confirmed diagnosis of infection. The purpose of the guidelines is to prevent progression to severe forms of these diseases and the fatal events they may cause. The recommendations are intended for health professionals, including general, resident, and specialist physicians, nursing professionals, and medical and nursing students, who participate in caring for patients with suspected dengue, chikungunya, or Zika. They are also intended for health unit managers and the executive teams of national arboviral disease prevention and control programs, who are responsible for facilitating the process of implementing these guidelines.


Subject(s)
Humans , Male , Female , Pregnancy , Child , Adolescent , Adult , Middle Aged , Arbovirus Infections/diagnosis , Fluid Therapy/standards , Arbovirus Infections/drug therapy , Steroids/therapeutic use , Histamine Antagonists/therapeutic use
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