scholarly journals Generation and Characterization of Monoclonal Antibodies against Dengue Virus Type 1 for Epitope Mapping and Serological Detection by Epitope-Based Peptide Antigens

2007 ◽  
Vol 14 (4) ◽  
pp. 404-411 ◽  
Author(s):  
Yun-Ching Chen ◽  
Hsien-Neng Huang ◽  
Chin-Tarng Lin ◽  
Yi-Fang Chen ◽  
Chwan-Chuen King ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Dengue Virus ◽  
Public Health Problem ◽  
Hemorrhagic Fever ◽  
Mouse Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Random Peptide Library ◽  
Peptide Antigens ◽  
Neutralizing Epitopes

ABSTRACT Dengue virus (DEN), the pathogen behind dengue hemorrhagic fever, remains a public health problem in Asia and South America. In this study, monoclonal antibodies (MAbs) against DEN serotype 1 (DEN-1) were generated by fusing NSI/1-Ag4-1 mouse myeloma cells with lymphocytes from BALB/c mice immunized with DEN-1. Twelve MAbs were found to react specifically to the DENs by enzyme-linked immunosorbent assay, immunofluorescence analysis, and immunoblotting analysis. Five MAbs, namely, DA4-7, DA6-7, DA9-5, DA10-2, and DA11-13, were found to react with envelope proteins of DEN-1. Two serotype-specific MAbs of DEN-1, DA6-7 and DA11-13, were further shown to neutralize DEN-1 infection by a plaque reduction neutralization test. The neutralizing epitopes of these MAbs were further identified from a random peptide library displayed on phage. Immunopositive phage clones reacted specifically with these MAbs and did not react with normal mouse serum. Epitope-based peptide antigens were proved able to detect antibodies in serum samples collected from DEN-1-infected patients but not in those taken from DEN-2-infected patients or healthy controls. We believe that these MAbs and neutralizing epitopes will provide information that will lead to the development of DEN-1 serotype-specific diagnostic reagents and vaccines.

scholarly journals Serological and Molecular Study of Dengue Viruses in Different Hospitals of Nepal

2013 ◽  
Vol 2 (1) ◽  
pp. 20-25 ◽  
Author(s):  
GP Gupta ◽  
Y Shah ◽  
A Poudel ◽  
R Pun ◽  
KP Pant ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Public Health Problem ◽  
Enzyme Linked Immunosorbent Assay ◽  
Age Group ◽  
Serum Samples ◽  
Important Public Health Problem ◽  
Monsoon Period ◽  
Occupation Group ◽  
Hilly Region ◽  
Acute Patients

Background: Dengue Virus (DV) is an emerging mosquito borne viral disease and important public health problem in low land of Terai region which is also expanding to hilly region. Methods: This study was designed to estimate sero-prevalence of dengue virus infection in the post monsoon period (Jun-Dec) of 2010 in Nepalese patients with fever visiting hospitals of Birganj, Damouli, Biratnagar and Dhading Besi. Serum samples were collected from 280 patients visiting hospitals with history of fever & clinically suspected dengue fever. The sero-prevalence of dengue virus specific IgM was determined by enzyme linked immunosorbent assay (ELISA) kit (SD, Korea) Results: The anti-dengue IgM positivity was found to be 8.2%. The positive dengue cases were higher in male (10.5%) as compared to female (6.5%). Among different age groups, the highest positive cases (11.5 %) were from age group below 15 years followed by above 50 years age group with 8.5%. Out of 4 hospitals, the highest positive cases were in Tanahu District Hospital, Damouli (23.8%) followed by Koshi Zonal Hospital, Biratnagar (12.5%). Age and gender were found to be independent predictors. The highest numbers of dengue positive cases were in occupation group business (13.3%) followed by agriculture (11.5%). Conclusion: The dengue positivity was estimated in acute patients from different hospitals of Nepal by enzyme immunoassay and reverse transcriptase polymerase chain reaction. Therefore, the serological marker can be used to diagnose the acute patients of dengue during outbreaks. Nepal Journal of Medical Sciences | Volume 02 | Number 01 | Jan-Jun 2013 | Page 20-25 DOI: http://dx.doi.org/10.3126/njms.v2i1.7646

scholarly journals Dengue Virus Detection by Serological and Molecular Method in Different Hospitals of Nepal

2013 ◽  
Vol 11 (2) ◽  
pp. 24-28
Author(s):  
Yogendra Shah ◽  
Govind Prasad Gupta ◽  
Kishor Pandey ◽  
Sher Bahadur Pun ◽  
Krishna Prasad Pant ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Age Groups ◽  
Virus Detection ◽  
Public Health Problem ◽  
Viral Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Age Group ◽  
Rt Pcr ◽  
Serum Samples ◽  
Disease Hospital

Introduction: Dengue is an emerging mosquito-borne viral disease in the world and is the serious public health problem of Nepal. Methods: This study was designed to determine sero-epidemiology of dengue virus infection during the period (June-Nov) of 2010 among suspected patients with fever visiting Koshi Zonal Hospital (KZH), Biratnagar, Narayani sub-regional Hospital (NSH), Birgunj, Sukraraj Tropical and Infectious Disease Hospital (STIDH), Kathmandu and Dhading District Hospital (DDH), Dhadingbeshi. The sero-prevalence of anti-dengue IgM antibody was determined by enzyme linked immunosorbent assay (ELISA). Results: Among 271 serum samples tested, the anti-dengue IgM positivity was 14.4%. Sero-positivity in male was 10.7% of total and that in female was 3.7%.  Among different age groups, the highest positive cases 11.8% were from age group 15-50 years and found least among the age group above 50 years 0.4%. Out of 4 different hospitals, the highest positive positive cases from STIDH with 9.2% and the least positive cases were from DDH (0.4%). RT-PCR showed 4.7% positivity of 21 samples tested. Conclusions: Enzyme immunoassay and RT-PCR serological marker can be used to diagnose the acute patients of dengue during outbreaks.Medical Journal of Shree Birendra Hospital; July-December 2012/vol.11/Issue2/24-27 DOI: http://dx.doi.org/10.3126/mjsbh.v11i2.7905 

Difusi dan Pola Spasial Sebaran Penyakit Demam Berdarah Dengue (DBD) Di Kota Bandar Lampung

2018 ◽  
Vol 1 (1) ◽  
pp. 87-95
Author(s):  
Nurul Qamila ◽  
Agel Vidian Krama
Keyword(s):  
Public Health ◽  
Spatial Analysis ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Spatial Pattern ◽  
Dengue Hemorrhagic Fever ◽  
Health Problem ◽  
Public Health Problem ◽  
Hemorrhagic Fever ◽  
Descriptive Method

Dengue hemorrhagic fever (DHF) is a contagious disease caused by the dengue virus and is transmitted by the mosquito Aedes aegypti (Aa.aegypti). The population is still a public health problem that increases the number of sufferers and also widespread, with population and education. This study aims to reveal the spatial pattern and distribution of Dengue Hemorrhagic Fever (DHF) with the spatial pattern and the spread of Dengue Hemorrhagic Fever (DHF) can result in different locations of these allegations. From the map that can be used for the prevention of Dengue Hemorrhagic Fever (DBD) in Bandar Lampung City. This study aims to reveal the spatial pattern and distribution of Dengue Hemorrhagic Fever (DHF) with the descriptive method and spatial pattern of Dengue Hemorrhagic Fever (DHF) can result in different locations of these allegations. From the map that can be used for the prevention of Dengue Hemorrhagic Fever (DBD) in Bandar Lampung City. Keywords: DHF, Spatial Analysis

scholarly journals Diagnosis of Dengue Virus Infection by Detection of Specific Immunoglobulin M (IgM) and IgA Antibodies in Serum and Saliva

2003 ◽  
Vol 10 (2) ◽  
pp. 317-322 ◽  
Author(s):  
Angel Balmaseda ◽  
María G. Guzmán ◽  
Samantha Hammond ◽  
Guillermo Robleto ◽  
Carolina Flores ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Immunoglobulin M ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Serum Samples ◽  
Diagnostic Modality ◽  
Iga Antibodies ◽  
Specific Immunoglobulin ◽  
Feasible Alternative

ABSTRACT To evaluate alternative approaches to the serological diagnosis of dengue virus (DEN) infection, the detection of DEN-specific immunoglobulin M (IgM) and IgA antibodies in serum and saliva specimens was assessed in 147 patients with symptoms of DEN infection seen at the Ministry of Health in Nicaragua. Seventy-two serum samples were determined to be positive for anti-DEN antibodies by IgM capture enzyme-linked immunosorbent assay, the routine diagnostic procedure. Serum and saliva specimens were obtained from 50 healthy adults as additional controls. IgM was detected in the saliva of 65 of the 72 serum IgM-positive cases, 6 of the 75 serum IgM-negative cases, and none of the control group, resulting in a sensitivity of 90.3% and a specificity of 92.0% and demonstrating that salivary IgM is a useful diagnostic marker for DEN infection. Detection of IgA in serum may be another feasible alternative for the diagnosis of DEN infection, with serum IgA found in 68 (94.4%) of the IgM-positive cases. In contrast, detection of IgA in saliva was not found to be a useful tool for DEN diagnosis in the present study. Further studies of the kinetics of antibody detection in another set of 151 paired acute- and convalescent-phase serum samples showed that DEN-specific IgA antibodies were detected in more acute-phase samples than were IgM antibodies. Thus, we conclude that DEN-specific IgA in serum is a potential diagnostic target. Furthermore, given that saliva is a readily obtainable, noninvasive specimen, detection of DEN-specific salivary IgM should be considered a useful, cheaper diagnostic modality with similar sensitivity and specificity to IgM detection in serum.

scholarly journals Immunoglobulin A Antibody Responses in Dengue Patients: a Useful Marker for Serodiagnosis of Dengue Virus Infection

2005 ◽  
Vol 12 (10) ◽  
pp. 1235-1237 ◽  
Author(s):  
M. Nawa ◽  
T. Takasaki ◽  
M. Ito ◽  
S. Inoue ◽  
K. Morita ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Immunoglobulin A ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Serum Samples ◽  
Dengue Virus Infection ◽  
Iga Antibody ◽  
Antibody Capture

ABSTRACT We determined the usefulness of an immunoglobulin A (IgA) antibody-capture enzyme-linked immunosorbent assay for serodiagnosis of dengue virus infections. The results indicate that the presence of IgA and IgM in serum samples assures recent primary dengue virus infection even with a single serum sample.

scholarly journals An Antigen Capture Enzyme-Linked Immunosorbent Assay Reveals High Levels of the Dengue Virus Protein NS1 in the Sera of Infected Patients

2000 ◽  
Vol 38 (3) ◽  
pp. 1053-1057 ◽  
Author(s):  
Paul R. Young ◽  
Paige A. Hilditch ◽  
Cheryl Bletchly ◽  
Wendy Halloran
Keyword(s):  
Monoclonal Antibodies ◽  
Dengue Virus ◽  
Acute Phase ◽  
Immunosorbent Assay ◽  
Nonstructural Protein ◽  
Surrogate Marker ◽  
Detection Sensitivity ◽  
Severe Dengue ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay

We describe the development of a capture enzyme-linked immunosorbent assay for the detection of the dengue virus nonstructural protein NS1. The assay employs rabbit polyclonal and monoclonal antibodies as the capture and detection antibodies, respectively. Immunoaffinity-purified NS1 derived from dengue 2 virus-infected cells was used as a standard to establish a detection sensitivity of approximately 4 ng/ml for an assay employing monoclonal antibodies recognizing a dengue 2 serotype-specific epitope. A number of serotype cross-reactive monoclonal antibodies were also shown to be suitable probes for the detection of NS1 expressed by the remaining three dengue virus serotypes. Examination of clinical samples demonstrated that the assay was able to detect NS1 with minimal interference from serum components at the test dilutions routinely used, suggesting that it could form the basis of a useful additional diagnostic test for dengue virus infection. Furthermore, quantitation of NS1 levels in patient sera may prove to be a valuable surrogate marker for viremia. Surprisingly high levels of NS1, as much as 15 μg/ml, were found in acute-phase sera taken from some of the patients experiencing serologically confirmed dengue 2 virus secondary infections but was not detected in the convalescent sera of these patients. In contrast, NS1 could not be detected in either acute-phase or convalescent serum samples taken from patients with serologically confirmed primary infection. The presence of high levels of secreted NS1 in the sera of patients experiencing secondary dengue virus infections, and in the context of an anamnestic antibody response, suggests that NS1 may contribute significantly to the formation of the circulating immune complexes that are suspected to play an important role in the pathogenesis of severe dengue disease.

Correlation of serostatus and viraemia levels among Indian dengue patients at the time of first diagnosis

2020 ◽  
Vol 114 (7) ◽  
pp. 513-520
Author(s):  
Ruta Kulkarni ◽  
Shubham Shrivastava ◽  
Harshad P Patil ◽  
Divya Tiraki ◽  
Akhilesh Chandra Mishra ◽  
...  
Keyword(s):  
Public Health Problem ◽  
Vero Cells ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Markers ◽  
Serum Samples ◽  
Infectious Dose ◽  
Therapeutic Monoclonal Antibodies ◽  
Tissue Culture Infectious Dose ◽  
First Diagnosis

Abstract Background Dengue is a public health problem worldwide. Therapeutic monoclonal antibodies (MAbs) against dengue virus (DENV) are likely to be available soon. In view of the feasibility issues pertaining to pretreatment viraemia quantitation for therapy decisions, we conducted this study for investigation of a correlation between patient serostatus (NS1/immunoglobulin M [IgM]/IgG) and viraemia levels among Indian dengue patients at the time of first diagnosis. Methods The study included 297 serum samples from dengue patients in Pune, India. The samples were tested for NS1, IgM and IgG (capture enzyme-linked immunosorbent assay [ELISA] for identifying secondary dengue) using Panbio ELISAs. Quantitation of viraemia was conducted using an NS1 ELISA-based 50% tissue culture infectious dose (TCID50) test in Vero cells. Results Viraemia was detectable only among NS1-positive patients (n = 229, range 0.5–8.3 logTCID50/ml) with a mean titre of 1.9 logTCID50/ml. Among the NS1-positive patients, DENV titres were higher in IgM-negative than IgM-positive patients (p < 0.0001) and in primary (IgG < 18 Panbio units) versus secondary (IgG > 22 Panbio units) dengue patients (p = 0.002). Virus titres were higher during the first 3 days of illness and decreased later (p = 0.005). Conclusions The study provides a range of infectious DENV titres in relation to serologic status among dengue patients in India. The data suggest the possibility of using serological markers (NS1/IgM) as a basis for treatment decisions.

scholarly journals Performance Evaluation of Commercial Dengue Diagnostic Tests for Early Detection of Dengue in Clinical Samples

2017 ◽  
Vol 2017 ◽  
pp. 1-4 ◽  
Author(s):  
Tuan Nur Akmalina Mat Jusoh ◽  
Rafidah Hanim Shueb
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Diagnostic Test ◽  
Rapid Diagnostic Test ◽  
Dengue Infection ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Kappa Agreement

The shattering rise in dengue virus infections globally has created a need for an accurate and validated rapid diagnostic test for this virus. Rapid diagnostic test (RDT) and reverse transcription-polymerase chain reaction (RT-PCR) diagnostic detection are useful tools for diagnosis of early dengue infection. We prospectively evaluated the diagnostic performance of nonstructural 1 (NS1) RDT and real-time RT-PCR diagnostic kits in 86 patient serum samples. Thirty-six samples were positive for dengue NS1 antigen while the remaining 50 were negative when tested with enzyme-linked immunosorbent assay (ELISA). Commercially available RDTs for NS1 detection, RTK ProDetect™, and SD Bioline showed high sensitivity of 94% and 89%, respectively, compared with ELISA. GenoAmp® Trioplex Real-Time RT-PCR and RealStar® Dengue RT-PCR tests presented a comparable kappa agreement with 0.722. The result obtained from GenoAmp® Real-Time RT-PCR Dengue test showed that 14 samples harbored dengue virus type 1 (DENV-1), 8 samples harbored DENV-2, 2 samples harbored DENV-3, and 1 sample harbored DENV-4. 1 sample had a double infection with DENV-1 and DENV-2. The NS1 RDTs and real-time RT-PCR tests were found to be a useful diagnostic for early and rapid diagnosis of acute dengue and an excellent surveillance tool in our battle against dengue.

scholarly journals Sero-diagnosis of Dengue virus in Different Hospitals of Nepal

2013 ◽  
Vol 1 (2) ◽  
pp. 58-62 ◽  
Author(s):  
Y Shah ◽  
G Khadka ◽  
GP Gupta ◽  
N Adhikari ◽  
A Poudel ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Dengue Fever ◽  
Control Measure ◽  
Public Health Problem ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dengue Virus Infection ◽  
Capture Elisa ◽  
Occupation Group ◽  
Hilly Region

INTRODUCTION: Dengue fever (DF) is an emerging mosquito borne viral disease and important public health problem in low land Terai region which is also moving towards hilly region Nepal. This study was designed to determine the sero-prevalence of dengue virus infection in patients visiting hospitals of Nepal. MATERIALS AND METHODS: This study was conducted during period (June-November) of 2010 in Nepalese patients with fever visiting hospitals of Birganj, Damouli, Biratanagar, Dhading Besi and Chitwan. The sero-prevalence of dengue virus specific IgM was determined by enzyme linked immunosorbent assay (ELISA). Serum samples were collected from 289 patients visiting hospitals with history of fever and clinically suspected dengue fever. RESULTS: The anti-dengue IgM positivity was found to be 8.99%. The positive dengue cases were higher in male (10.8%) as compared to female (7.1%) though it was not statistically significant (P>0.05). Among different age groups, the highest positive cases (12.3%) were from age group below 15 years followed by above 50 years 8.3%. Out of 5 hospitals, the highest positive cases were in Tanahu hospital, Damouli (23.8%) followed by Bharatpur hospital and Chitwan (22.2%). Age and gender were found to be independent predictors. The highest numbers of dengue positive cases were in occupation group business (13.3%) followed by agriculture (12.7%). CONCLUSIONS: Prevalence of dengue virus infection is increasing and proper control measure should be provided. IgM capture ELISA was used for laboratory analysis and remains as a reliable and inexpensive method for the diagnosis of dengue. Hence, the IgM capture ELISA has become the most accepted technique for the diagnosis of dengue in developing countries like Nepal. DOI: http://dx.doi.org/10.3126/ijim.v1i2.7003 Int J Infect Microbiol 2012;1(1):58-62

scholarly journals Prospective Study To Determine Accuracy of Rapid Serological Assays for Diagnosis of Acute Dengue Virus Infection in Laos

2007 ◽  
Vol 14 (11) ◽  
pp. 1458-1464 ◽  
Author(s):  
Stuart D. Blacksell ◽  
David Bell ◽  
James Kelley ◽  
Mammen P. Mammen ◽  
Robert V. Gibbons ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Patient Management ◽  
Limited Resource ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Serum Samples ◽  
Dengue Virus Infection ◽  
Resource Setting

ABSTRACT There is an urgent need for accurate and simple dengue virus infection diagnostic assays in limited-resource settings of dengue endemicity, to assist patient management. Using a panel of reference samples (S. D. Blacksell, P. N. Newton, D. Bell, J. Kelley, M. P. Mammen, D. W. Vaughn, V. Wuthiekanun, A. Sungkakum, A. Nisalak, and N. P. Day, Clin. Infect. Dis. 42:1127-1134, 2006), we recently evaluated eihgt commercially available immunochromatographic rapid diagnostic tests (RDTs) designed to detect dengue virus-specific immunoglobulin M (IgM) and/or IgG. We found that 6/8 RDTs had sensitivities of less than 50% (range, 6 to 65%), but specificities were generally high. Here, in conjuction with dengue virus serotyping by reverse transcriptase PCR and in the limited-resource setting of Laos, where dengue virus is endemic, we evaluated the same eight RDTs against a previously validated dengue IgM/IgG enzyme-linked immunosorbent assay for diagnosis of acute dengue virus infection. Paired serum samples were collected from 87 patients, of whom 38 had confirmed dengue virus infections (4 had primary infections, 33 had secondary infections, and 1 had an infection of indeterminate status). RDT sensitivity was low, with 7/8 RDTs having admission sample sensitivities of less than 20% (range, 4 to 26%). The majority (6/8) of the RDTs, demonstrated high specificity (>95%). Kappa statistic values ranged from 6 to 54% for the RDTs, demonstrating poor to moderate variation between three operators. No RDT adequately differentiated between primary and secondary dengue virus infections. The findings of this study suggest that currently available RDTs based on the detection of IgM antibodies for the diagnosis of acute dengue virus infections are unlikely to be useful for patient management.

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