The use of two-dimension electrophoresis to identify serum biomarkers from patients with dengue haemorrhagic fever (original) (raw)
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Discovery and characterization of potential prognostic biomarkers for dengue hemorrhagic fever
The American journal of tropical medicine and hygiene, 2014
Half a million patients are hospitalized with severe dengue every year, many of whom would die without timely, appropriate clinical intervention. The majority of dengue cases are uncomplicated; however, 2-5% progress to severe dengue. Severe dengue cases have been reported with increasing frequency over the last 30 years. To discover biomarkers for severe dengue, we used surface-enhanced laser desorption/ionization time-of-flight mass spectrometry to analyze dengue virus positive serum samples from the acute phase of infection. Using this method, 16 proteins were identified as candidate biomarkers for severe dengue. From these 16 biomarkers, three candidates were selected for confirmation by enzyme-linked immunosorbent assay and Western blot: vitronectin (Vtn, 55.1 kDa), hemopexin (Hx, 52.4 kDa), and serotransferrin (Tf, 79.2 kDa). Vitronectin, Hx, and Tf best differentiated between dengue and severe dengue.
Diagnosis of Acute Dengue Virus Infection Using Enzyme-Linked Immunosorbent Assay and Real-Time PCR
Canadian Journal of Infectious Diseases & Medical Microbiology, 2023
Dengue fever is a viral infection caused by the dengue virus and is a growing concern for public health worldwide, particularly in tropical and subtropical regions. Tis study aimed to assess the diagnostic accuracy of a commercially available NS1 ELISA kit for dengue fever in Pakistan using multiplex qRT-PCR as the gold standard. Te study recruited 1236 suspected cases of dengue fever admitted to public sector hospitals in Lahore, Pakistan. Of the suspected cases, 610 (49.3%) were confrmed positive for DENV infection through qRT-PCR, with all four serotypes detected. DENV-2 was the most prevalent serotype, detected in 95.7% of cases. Te NS1 ELISA kit detected 71.1% of the positive cases. However, the diagnostic accuracy of the NS1 ELISA kit was found to be only 64.89%. Of the 610 confrmed cases, 68% were male and 32% were female, with a median age of 30 years. Dengue fever was diagnosed in 91.8% of cases, while 8.2% were diagnosed with dengue hemorrhagic fever (DHF). DHF patients had a higher prevalence of abdominal pain, hemorrhagic manifestations, and thrombocytopenia. Te cocirculation of all four DENV serotypes in Lahore is concerning and could lead to more severe forms of the disease, such as DHF or dengue shock syndrome, in the future. Te study highlights the low diagnostic accuracy of commercially available NS1 ELISA kits and emphasizes the importance of using molecular methods to confrm acute dengue infections. Given the increasing prevalence of dengue fever in developing countries like Pakistan, more accurate and reliable diagnostic tools are needed for efective disease management and control.
Issues in contemporary and potential future molecular diagnostics for dengue
Expert Review of Molecular Diagnostics, 2016
Introduction: Dengue has been the most common arbovirus infection worldwide with 2.5 billion people living in over 100 endemic tropical and subtropical regions. Due to the high number of asymptomatic cases and the signs and symptoms being rather unspecific, dengue cases are often under-reported and might influence dengue surveillance programs. Therefore, a rapid, easy to use, inexpensive, and highly sensitive and specific diagnostic tool is essential for early and accurate diagnosis to ease the clinical management of patients as well as for the development of new interventions. Areas covered: This report discusses the contemporary dengue diagnostic tool, mainly from the aspect of molecular diagnosis where an overview of several nuclei acid amplification tests has been included. Potential molecular diagnostic tools such as biosensor and microarray are also discussed in this report. Expert commentary: Rapidness and accuracy in terms of sensitivity and specificity is imperative in dengue diagnosis for both clinical management and surveillance of dengue to ensure early treatment and corrective control measures can be carried out. In the next five years it is expected that there will be newer tests developed using not only the lateral flow techniques but more specifically biosensors and nanotechnology. These new technologies will have to be validated with the appropriate number and category of samples and to address the issue of cross-reactivity.
Early molecular markers predictive of dengue hemorrhagic fever
Anais da Academia Brasileira de Ciências, 2009
The management of acute dengue patients during outbreaks is a challenging problem. Most of the dengue fever cases are benign, but some cases develop into a severe and possibly lethal vasculopathy, known as dengue hemorrhagic fever. Early symptoms of dengue and hemorrhagic fever are very similar. An early differential diagnosis is needed to predict which of these two clinical presentations is crucial to proper patient care and public health management. This study evaluates the predictive potential of specific mRNA expression markers of dengue hemorrhagic fever using quantitative real-time PCR assays. Six candidate "dengue hemorrhagic fever specific signature genes" were evaluated and all showed good correlation among their transcription levels at early days of infection and the later development of severe vasculopathy. The markers selected were able to indicate, at early stages of infection, the evolution of a dengue-infected patient to the severe form of the illness. Despite the fact that these results grant further validation studies, the panel of candidate prognostic markers obtained demonstrated the potential to be useful for clinical use in the form of a fast assay based in blood samples. 81 (4) 672 CARLOS E. CALZAVARA-SILVA et al.
Characterization of Dengue Infections by Using Hematological and Serological Markers
2020
Methods: Two groups were selected by non-probability random sampling technique including case group of 154 patients with suspected dengue (fever>2days and <10days) and control group of 146 patients with febrile illness other than dengue. Clinical, hematological and serologic markers of cases and control groups were analyzed. The frequency distribution was used to compare categorical serologic markers and paired sample T test was applied for hematologic variables before and after treatment of dengue using SPSS version 21. Results: A total of 154 cases of dengue (by using WHO calculator 154 suspected cases were defi ned fever >2 days and < 10days having hematologic and NS 1positive in earlier phase of disease) and 146 control cases were observed. Serology markers using NS1 dengue were detected positive in (85/154) 55%, dengue IgM in (46/154) 30% and both NS1 and dengue IgM in 15%(23/154) cases. Control group included malaria, enteric fever and febrile illness 103(71%), 30(20%), 13(9%) respectively. In cases 70(45%) were females and 85(55%) males with signifi cant p value, while in control group 80(55%) were females and 66(45%) males with p value of < 0.05. For Hematological markers including platelets count, hematocrit and WBCs, paired sample T test showed high signifi cant diff erence in value of platelets counts and hematocrit , while no signifi cant diff erence in WBCS was observed. Conclusion: Serological markers (NS1, IgM and IgG) greatly helps in diagnosis as well as prognosis of dengue infection, thereby helping not only in reducing the morbidity and mortality in patients but also helps controlling the epidemic by observing early preventive measures by using Hematological markers in earlier phase of disease. These marker will make utilized by other Asian and tropical African countries.
Medicine, 2018
Dengue virus is one of the most widespread flaviviruses that re-emerged throughout recent decades. The progression from mild dengue to severe dengue (SD) with the complications such as vascular leakage and hemorrhage increases the fatality rate of dengue. The pathophysiology of SD is not entirely clear. To investigate potential biomarkers that are suggestive of pathogenesis of SD, a small panel of serum samples selected from 1 healthy individual, 2 dengue patients without warning signs (DWSÀ), 2 dengue patients with warning signs (DWS+), and 5 patients with SD were subjected to a pilot analysis using Sengenics Immunome protein array. The overall fold changes of protein expressions and clustering heat map revealed that PFKFB4, TPM1, PDCL3, and PTPN20A were elevated among patients with SD. Differential expression analysis identified that 29 proteins were differentially elevated greater than 2fold in SD groups than DWSÀ and DWS+. From the 29 candidate proteins, pathways enrichment analysis also identified insulin signaling and cytoskeleton pathways were involved in SD, suggesting that the insulin pathway may play a pivotal role in the pathogenesis of SD. Abbreviations: AFF4 = AF4/FMR2 Family Member 4, BCCP = biotin carboxyl carrier protein, COPS6 = COP9 Signalosome Subunit 6, CV = coefficient of variant, Cy3BSA = Cy3 labelled bovine serum albumin, DAMPs = damage-associated molecular patterns, DENV = dengue virus, DWSÀ = dengue patients without warning signs, DWSÀ = dengue warning signs negative (dengue with warning signs), DWS+ = dengue patients with warning signs, DWS+ = dengue warning signs positive (dengue without warning signs), ELISA = enzyme-linked immunosorbent assay, FC = fold change, HC = healthy control, HI = hemagglutination inhibition, KRT19 = Keratin 19, NS1 = nonstructural protein 1, PCR = polymerase chain reaction, PDCL3 = phosducin like 3, PFKFB4 = 6phosphofructo-2-kinase/ fructose-2 =6-biphosphatase 4, PTPN20A = protein tyrosine phosphatase; nonreceptor type 20A, RFU = relative fluorescence unit, SAB = serum assay buffer, SD = patients with severe dengue, SD = severe dengue, TP53 = tumor protein p53, TPM1 = tropomyosin-alpha 1, VIAF = viral IAP-associated factor 1.
Title A Proteomic Approach Identifies Candidate Early Biomarkers to PredictSevere Dengue in Children
2018
Background Severe dengue with severe plasma leakage (SD-SPL) is the most frequent of dengue severe form. Plasma biomarkers for early predictive diagnosis of SD-SPL are required in the primary clinics for the prevention of dengue death. Methodology Among 63 confirmed dengue pediatric patients recruited, hospital based longitudinal study detected six SD-SPL and ten dengue with warning sign (DWS). To identify the specific proteins increased or decreased in the SD-SPL plasma obtained 6-48 hours before the shock compared with the DWS, the isobaric tags for relative and absolute quantification (iTRAQ) technology was performed using four patients each group. Validation was undertaken in 6 SD-SPL and 10 DWS patients. Principal findings Nineteen plasma proteins exhibited significantly different relative concentrations (p<0.05), with five over-expressed and fourteen under-expressed in SD-SPL compared with DWS. The individual protein was classified to either blood coagulation, vascular regulation, cellular transport-related processes or immune response. The immunoblot quantification showed angiotensinogen and antithrombin III significantly increased in SD-SPL whole
2018
Background: Dengue is a serious, rapidly spreading tropical mosquito-borne viral disease caused by dengue virus belonging to the family flaviviridae and genus flavivirus in the last five decades. In many dengue endemic remote areas with limited laboratory diagnostic services their rapid diagnostic tests are simple method which provides the immediate diagnosis and early management. Materials and methods: The study includes clinically suspected 1236 cases with all haematological parameters and tested at clinical laboratory in which 500 serum samples showed positivity for one or more markers like NS1, IgM or IgG antibodies. Results: In our study among 500 seropositive cases thrombocytopenia (<1 lakh) was evident in 66.7% and in 736 seronegative cases thrombocytopenia was seen in 27.47%. Out of 323 NS1 positive cases, thrombocytopenia was evident in 56.0% and in exclusive antibody positive cases thrombocytopenia was noted in 19.2%. These associations were considered to be statistical...