Anti-dengue Antibodies Research Articles

Epidemiology and immuno-molecular status of Nepal dengue outbreak

Background: The emergence and circulation of dengue infection have become one of the major public health concerns in the world with its high prevalence in the sub-tropical and tropical countries. Hundreds of Dengue cases have been reported annually and the shift of serotypes in every year has raised the concern of public health in Nepal. The detection of dengue virus infections and circulation of different serotypes in different areas has a great role for the clinical management of patients, surveillance, and clinical trial assessments in the days to come. Methods and materials: Clinically suspected dengue cases (n = 240) were subjected to sero analysis for anti-dengue IgM, IgG antibody and NS1 antigen followed by IgG end-point ELISA. Viral RNA extraction was performed for all samples using the Qiagen kit. cDNA was prepared, and RT-PCR was performed using D1 and DencomR2 primers. Dengue Serotyping was done for PCR positive samples using gene-specific primers. Results: Out of 240 dengue suspected cases, male (n = 153/63.75%), female (n = 87/36.25%) and children below 10 years (6.25%) were diagnosed as positive by ELISA. NS1 (59.16%), IgM (33%) and IgG (7.9%) were detected by ELISA, however, only 14 samples were positive alone to each test. Most of the cases were found to be of primary infection. Antibody level for each serum was calculated and categorized as high (43228.70), medium and low (953.53). Only 35.83% of the suspected cases were confirmed positive for dengue virus by PCR and were of Dengue Serotype-1 in the year 2016 in Nepal. Conclusion: The single test alone could not be confirmatory as only 35.83% of the clinically suspected cases were confirmed for Dengue virus infection by PCR. This implies that confirmatory tests like PCR should be coupled with the ELISA results for the proper diagnosis and management of the disease.

Performance of Dried Blood Spots Compared with Serum Samples for Measuring Dengue Seroprevalence in a Cohort of Children in Cebu, Philippines.

.Dengue seroprevalence data are useful for understanding epidemiologic trends and transmission dynamics, and for making decisions about implementation of dengue control programs. A logistical challenge to seroprevalence surveys is the collection and transport of serum samples. For conducting large and repeated dengue serosurveys, dried blood spots (DBS) would allow easier sample collection, shipment, transport, and storage than standard serum collection methods. Further evidence is needed to understand how well DBS performs compared with standard serum collection methods in laboratory assays. We evaluated the detection of anti-dengue antibodies by IgG indirect ELISA when using DBS compared with sera. Specimens were collected from healthy children in Cebu, Philippines, who would be 9–14 years of age at the time of a mass dengue vaccination program. Using an ELISA index value cutoff of 0.9, 1,285/1,488 (86.4%) of the DBS were seropositive and 203 (13.6%) were seronegative, compared with 1,292/1,488 (86.8%) seropositive and 196 (13.2%) seronegative serum samples. Compared with sera, the DBS method had a 98.3% sensitivity, 92.4% specificity, 98.9% positive predictive value, and 89.2% negative predictive value. Considering the advantages in terms of sample collection, shipment, and storage, DBS sampling may be appropriate for dengue population serosurveys.

Association of serum C-reactive protein level and polymorphisms with susceptibility to dengue infection and severe clinical outcome among eastern Indian patients.

Dengue virus (DENV) infection is a major public health concern in India ranging from simple febrile illness to severe outcome. This study aimed to investigate association of serum CRP level and CRP gene polymorphisms towards development of dengue disease susceptibility and severity among eastern Indian patients. Blood was collected from 348 symptomatic patients. Sera was subjected to serological diagnosis for the presence of anti-dengue IgM, anti-dengue IgG antibodies and dengue NS1 antigen by ELISA. Viral RNA was extracted and the presence of DENV genome, viral load, serotypes was determined by qRT-PCR. CRP level and polymorphisms were determined by immunoturbidimetry and polymerase chain reaction-restriction fragment length polymorphism, respectively. Statistical analysis was performed by GraphPad-Prism. Among 206 dengue patients, CRP level increased significantly among patients within acute phase, and patients with qRT-PCR/NS1 antigen positivity, high viral load (HVL), secondary infection, and DENV4 and DENV2 infections. rs3091244, TT genotype positively associated with dengue susceptibility (p = 0.03). CT genotype of rs3093059 and TT genotype of rs3091244 were found to correlate with elevated CRP level and development of WHO-defined warning signs. TT genotype of rs3091244 was more prevalent among HVL patients. Thus, these CRP polymorphic variants and CRP concentration might act as potential prognostic biomarkers for predicting disease severity among acute-stage dengue patients.

Dengue in parts of the Guinea Savannah region of Nigeria and the risk of increased transmission.

BackgroundDengue virus (DENV) is spreading to parts of the world where it had not been previously reported. Increased international travel has led to intercontinental importation of dengue by visitors returning from countries in sub-Saharan Africa. Although dengue is well documented in the rain forest region of Nigeria, there is a dearth of information in the Guinea Savannah region, which is a major transit point for local and international visitors in the most populous nation in Africa. We provide preliminary evidence of dengue activity in the Guinea Savannah and highlight the risk factors.MethodsBlood was collected from a cross-section of 200 patients attending four hospitals in Anyigba, Kogi State. Anti-dengue antibody was identified using DENV immunoglobulin G (IgG) immunoassays. Questionnaires were used to obtain sociodemographic variables and risk factors. Data were analysed with SPSS version 16.0 for Windows.ResultsForty-two (20.5%) participants had anti-DENV IgG antibodies. Persons within the 45–59 y age group were more seropositive, with a rate of 35%. Males were more seropositive compared with females. Marriage, formal education, involvement in business activities and the presence of grasses around homes were associated with higher IgG seropositivity. The presence of open water containers around human dwellings and a lack of mosquito net use are predisposing factors.ConclusionsThis study identified past exposure to DENV among people in Anyigba, located in the Guinea Savannah region. Proper diagnosis of febrile episodes is required to improve case management and curtail off-target treatment. The high rate of previous exposure of patients to dengue indicates the need to strengthen vector control and dengue surveillance programs.

A STUDY TO DETERMINE THE INCREASING DENGUE INCIDENCE IN BIHAR REGION: AN OBSERVATIONAL STUDY.

Introduction: Dengue is one of the most serious mosquitos borne viral infection mainly affecting tropical and subtropical countries of the world. In absence of specific treatment and vaccine for dengue fever (DF); vector control is the only method by which spread of dengue can be prevented. The present study was conducted to determine the increasing dengue incidence in Bihar region.
 Material and Method: This study was done to report the sero prevalence of Dengue virus infection at Bihar, North India. The laboratory records of clinically suspected Dengue patients from jan 2018 to Nov 2018 were analyzed retrospectively for demographic features, seasonal variations, and results of IgM and IgG anti dengue antibodies tested by Dengue IGM capture enzyme linked immune sorbent assay (MAC ELISA).
 Results: A total of 1035 serum samples were analyzed. Out of which 209 samples (20.19%) were found positive for dengue virus infection. Maximum positive cases were seen in 2019 (53.11%). Seasonal trend showed that infection started appearing in august, peaked in October and slowly tapered by December.
 Conclusion: The most affected age group was 11 to 15 years of age, (57.3%) followed by 6 to10 year’s group (27.3 %)and least effective age group was 0 to 5 year (15.4%), majority of cases were found to be of secondary dengue virus infection (93.8%). Male to female ratio was 2:1.The present outbreak thus emphasizes the need for continuous sero epidemiological surveillance for the timely formulation and implementation of effective dengue control programme.
 Keywords: Dengue, dengue haemorrhagic fever, IgM antibody capture enzyme linked immune sorbent assay (MAC ELISA), India, vector.

Dengue NS1 detection in pediatric serum using microfluidic paper-based analytical devices.

The diagnosis of dengue infection is still a critical factor determining success in the clinical management and treatment of patients. Here, the development of microfluidic paper-based analytical devices (μPADs) utilizing a sandwich immunoassay on wax patterned paper functionalized with anti-dengue NS1 monoclonal antibodies for point-of-care detection of dengue NS1 (DEN-NS1-PAD) is reported. Various assay conditions, including the length of the channel and diluent, were optimized, and the response detected by the naked eye and digitized images within 20-30min. The DEN-NS1-PAD was successfully tested in the field for detecting dengue NS1 in buffer, cell culture media, and human serum. The limit of detection (LoD) of the DEN-NS1-PAD obtained with the naked eye, scanner, and a smartphone camera was 200, 46.7, and 74.8ngmL-1, respectively. The repeatability, reproducibility, and stability of the DEN-NS1-PAD were also evaluated. High true specificity and sensitivity in the serum of pediatric patients were observed. These evaluation results confirm that the DEN-NS1-PAD can potentially be used in point-of-care dengue diagnostics, which can significantly impact on the spreading of mosquito-borne diseases, which are likely to become more prevalent with the effects of global warming. Graphical Abstract.

Frequency of dengue infection in febrile patients attended Dhaka Medical College Hospital during January to December, 2018

Introduction: Dengue is a major public health concern in our country. The alarming thing is that the seasonal trend of dengue infection is changing with time in Bangladesh due to climate change and unplanned urbanization. Our study was conducted to determine the frequency of dengue virus infection among the febrile patients in 2018 at Dhaka Medical College Hospital (DMCH) and to observe the seasonal trend.
 Materials and Methods: The study was carried out on 899 febrile patients attended in DMCH from January to December, 2018. Whole blood samples were collected and sera were tested for dengue NS1 antigen and anti-dengue IgM antibodies using commercial test kits (NS1 by OMC Healthcare (Pvt.) Ltd & IgM antibody by Omega Diagnostics Ltd.), respectively. All negative dengue cases were tested for anti Chikungunya antibody to exclude chikungunya.
 Results: Of the total 899 febrile patients, 350(38.93%) were positive for Dengue infection. Out of them 264(75.43%) were positive for NS1, 82 (23.43%) were positive for IgM and 4 (1.14%) were positive for dengue NS1antigen + anti dengue IgM antibody. More than 50% patients belonged to age group 15-29 years. Males were predominant. More than 60% cases were detected in the post monsoon season.
 Conclusion: Highest dengue cases were detected in this year in comparison to the previous year’s probably due to re-emergence of DEN-3 serotype. Due to pattern of climate change, seasonal trend of dengue infection was not maintained. Moreover, unplanned urbanization and poor solid waste management have worsened the situation more.
 J Dhaka Medical College, Vol. 28, No.1, April, 2019, Page 105-111

An epidemiological survey of the current status of Zika and the immune interaction between dengue and Zika infection in Southern Taiwan

ObjectivesThis study was performed to examine the current status of Zika and the effects of pre-existing dengue immunity on Zika virus (ZIKV) infection in Southern Taiwan. MethodsA phylogenetic tree was used to analyze the phylogeny of detected ZIKVs. Paired sera from dengue patients were collected for the determination of dengue and Zika infection. Plaque reduction neutralization tests (PRNT) and quantitative reverse transcription PCR (qRT-PCR) were used to determine the titers of neutralizing antibodies and viruses, respectively. An antibody-dependent enhancement (ADE) assay was used to evaluate the effect of anti-dengue antibodies on ZIKV infection. ResultsEpidemiological data indicated the continuous importation of ZIKV infection from neighboring Zika epidemic countries into Taiwan. A total of 78 dengue patients were enrolled and 21 paired serum samples were obtained. PRNT90 results for the 21 samples identified eight cases of primary dengue infection and 13 cases of secondary dengue infection; two samples were positive for ZIKV (MR766). Results from the ADE assay indicated that convalescent sera from primary and secondary dengue infection patients displayed significant ADE of the ZIKV infection when compared to healthy controls (p < 0.05). ConclusionsThis study suggests that pre-existing dengue immunity facilitates ZIKV infection and that the continuous importation of ZIKV infection may pose a threat to indigenous Zika emergence in Southern Taiwan.

Towards Predicting Progression to Severe Dengue.

There is an urgent need for prognostic assays to predict progression to severe dengue infection, which is a major global threat. While the majority of symptomatic dengue patients experience an acute febrile illness, 5-20% progress to severe infection associated with significant morbidity and mortality. Early monitoring and administration of supportive care reduce mortality and clinically usable biomarkers to predict severe dengue are needed. Here, we review recent discoveries of gene sets, anti-dengue antibody properties, and inflammatory markers with potential utility as predictors of disease progression. Upon larger scale validation and development of affordable sample-to-answer technologies, some of these biomarkers may be utilized to develop the first prognostic assay for improving patient care and allocating healthcare resources more effectively in dengue endemic countries.

Transient Expression of Dengue Virus NS1 Antigen in Nicotiana benthamiana for Use as a Diagnostic Antigen

Dengue is a viral disease that represents a significant threat to global public health since billions of people are now at risk of infection by this mosquito-borne virus. The implementation of extensive screening tests is indispensable to control this disease, and the Dengue virus non-structural protein 1 (NS1) is a promising antigen for the serological diagnosis of dengue fever. Plant-based systems can be a safe and cost-effective alternative for the production of dengue virus antigens. In this work, two strategies to produce the dengue NS1 protein in Nicotiana benthamiana leaves were evaluated: Targeting NS1 to five different subcellular compartments to assess the best subcellular organelle for the expression and accumulation of NS1, and the addition of elastin-like polypeptide (ELP) or hydrophobin (HFBI) fusion tags to NS1. The transiently expressed proteins in N. benthamiana were quantified by Western blot analysis. The NS1 fused to ELP and targeted to the ER (NS1 ELP-ER) showed the highest yield (445 mg/kg), approximately a forty-fold increase in accumulation levels compared to the non-fused protein (NS1-ER), representing the first example of transient expression of DENV NS1 in plant. We also demonstrated that NS1 ELP-ER was successfully recognized by a monoclonal anti-dengue virus NS1 glycoprotein antibody, and by sera from dengue virus-infected patients. Interestingly, it was found that transient production of NS1-ER and NS1 ELP-ER using vacuum infiltration of whole plants, which is easier to scale up, rather than syringe infiltration of leaves, greatly improved the accumulation of NS1 proteins. The generated plant made NS1, even without extensive purification, showed potential to be used for the development of the NS1 diagnostic tests in resource-limited areas where dengue is endemic.

A Comprehensive Study of Dengue Epidemics and Persistence of Anti-Dengue Virus Antibodies in District Swat, Pakistan

Background: Dengue fever is one of the most common human arbovirus infections worldwide. In Pakistan, dengue initially became endemic in the big cities and then expanded to remote areas of the country. The current study reports the dengue epidemics, anti-DENV antibodies prevalence during the active and post-dengue infection, risk factors, disease symptoms, and spotting dengue infection densities in district Swat of Pakistan. Methods: Clinical signs and demographic data of dengue suspected individuals were collected at the time of screening through non-structural protein-1 antigen detection test during 2013–2015. Moreover, selected dengue confirmed individuals were screened for the presence of anti-dengue immunoglobulin (Ig) M and G during the active infection period and post-dengue infection. Results: A total of 8,770 individuals were infected with dengue in 2013 with 36 (0.41%) case fatalities, 307 in 2014 with no case fatality, and 13 in 2015 with no case fatality. The number of male and female cases were 6,139 and 2,631 in 2013, 183 and 124 in 2014, and only 10 and 3 in 2015, respectively. Among all the localities, Tehsil Babozai, an urban setting, reported the highest number of dengue patients during all the study years, that is, 7,673 (87.49% of the total cases) in 2013, 294 (95.76% of the total cases) in 2014, and 13 (100% cases) in 2015. Among 6 age groups, 21–30 years was found to be highly infected in 2013 (37.13% of all cases) and 2014 (33.55%). Furthermore, 1,231 (21.94% of all cases) had IgM antibodies and 71 (1.26%) had IgG antibodies in 2013, 78 (26% of all cases) had IgM antibodies and 7 (2.33%) had IgG antibodies in 2014, and only 4 (30.76%) patients had IgM and 0 (0%) had IgG antibodies in 2015. Furthermore, urban areas had the highest infection density in district Swat. The majority of the patients in rural areas had a traveling history to the urban areas before their illness. Conclusion: To sum up, male gender, young individuals, and those living in urban areas were at the greater risk of dengue infection.

Antibodies to Zika virus, Campylobacter jejuni, gangliosides in Guillain Barre syndrome. A prospective single centre study from southern India

The objective was to study the presence of antibodies against Zika virus, Camplyobacter jejuni and gangliosides in patients with Guillain Barre syndrome. Ninety consecutive patients (age more than 12 years) with GBS admitted to a tertiary care centre in southern India were included in this study. Data on the clinical manifestations, Nerve conduction studies and response to therapy were collected. The following tests were done in stored serum samples - Anti-Zika virus (IgM) antibodies, Anti-Campylobacter jejuni (IgG) antibodies and Anti-ganglioside antibodies (IgG). Those samples which were positive to Anti Zika antibodies were tested for conventional PCR for Zika virus, IgM antibodies against Dengue and Japanese encephalitis. Of the 90 patients, 3 died and 8 had persistent weakness. AIDP was the most common type of GBS (56.7%). Anti-ganglioside antibodies were present in 62.2% with GT1b being most common. Anti-C.jejuni antibodies were present in 46.6%. Anti-Zika antibodies (IgM) were present in 14 patients (15.5%). Four of these patients also had anti-Dengue antibody (IgM) positivity. This is one of the largest study on GBS from India and the first one to report on the presence of Zika antibodies form this geographical area. Our study had high prevalence of anti C.jejuni and anti ganglioside antibodies. Evidence of recent ZIKV infection as evidenced by anti IgM antibodies was present in 14 patients with 4 of them tested positive for anti-Dengue IgM antibody. Whether this represents cross reaction with Dengue or prior/co- infection with Dengue could not be addressed in this study.

Antibody-Dependent Dengue Virus Entry Modulates Cell Intrinsic Responses for Enhanced Infection

Dengue is caused by infection with any one of four dengue viruses (DENV); the risk of severe disease appears to be enhanced by the cross-reactive or subneutralizing levels of antibody from a prior DENV infection. These antibodies opsonize DENV entry through the activating Fc gamma receptors (FcγR), instead of infection through canonical receptor-mediated endocytosis, to result in higher levels of DENV replication. However, whether the enhanced replication is solely due to more efficient FcγR-mediated DENV entry or is also through FcγR-mediated alteration of the host transcriptome response to favor DENV infection remains unclear. Indeed, more efficient viral entry through activation of the FcγR can result in an increased viral antigenic load within target cells and confound direct comparisons of the host transcriptome response under antibody-dependent and antibody-independent conditions. Herein, we show that, despite controlling for the viral antigenic load in primary monocytes, the antibody-dependent and non-antibody-dependent routes of DENV entry induce transcriptome responses that are remarkably different. Notably, antibody-dependent DENV entry upregulated DENV host dependency factors associated with RNA splicing, mitochondrial respiratory chain complexes, and vesicle trafficking. Additionally, supporting findings from other studies, antibody-dependent DENV entry impeded the downregulation of ribosomal genes caused by canonical receptor-mediated endocytosis to increase viral translation. Collectively, our findings support the notion that antibody-dependent DENV entry alters host responses that support the viral life cycle and that host responses to DENV need to be defined in the context of its entry pathway.IMPORTANCE Dengue virus is the most prevalent mosquito-borne viral infection globally, resulting in variable manifestations ranging from asymptomatic viremia to life-threatening shock and multiorgan failure. Previous studies have indicated that the risk of severe dengue in humans can be increased by a specific range of preexisting anti-dengue virus antibody titers, a phenomenon termed antibody-dependent enhancement. There is hence a need to understand how antibodies augment dengue virus infection compared to the alternative canonical receptor-mediated viral entry route. Herein, we show that, besides facilitating viral uptake, antibody-mediated entry increases the expression of early host dependency factors to promote viral infection; these factors include RNA splicing, mitochondrial respiratory chain complexes, vesicle trafficking, and ribosomal genes. These findings will enhance our understanding of how differences in entry pathways can affect host responses and offer opportunities to design therapeutics that can specifically inhibit antibody-dependent enhancement of dengue virus infection.

Dengue and chikungunya seroprevalence among Qatari nationals and immigrants residing in Qatar.

The objective of this study is to characterize the seroprevalence of anti-dengue (DENV) and anti-chikungunya (CHIKV) antibodies among blood donors residing in Qatar who are Middle East and North Africa (MENA) nationals and non-nationals. Sera were collected from adult blood donors in Qatar from 2013 to 2016 and tested for anti-DENV and anti-CHIKV IgG using commercial microplate enzyme-linked immunosorbent assays. Age-specific seroprevalence was summarized by region/nationality: Asia (India, Philippines), Middle East (Iran, Jordan, Lebanon, Pakistan, Palestine, Syria, Yemen), North Africa (Egypt, Sudan), Qatar. The adjusted odds of anti-DENV and anti-CHIKV IgG seropositivity was estimated by logistic regression. Among 1,992 serum samples tested, Asian nationals had higher adjusted odds of being seropositive for anti-DENV antibodies compared to nationals of the Middle East (aOR 0.05, 95% CI 0.04–0.07), North Africa (aOR 0.14, 95% CI 0.10–0.20), and Qatar (aOR 0.01, 95% CI 0.01–0.03). Asian nationals also had higher adjusted odds of being seropositive for anti-CHIKV antibodies compared to those from the Middle East (aOR 0.14, 95% CI 0.07–0.27), North Africa (aOR 0.50, 95% CI 0.26–0.96), and Qatar (aOR 0.38, 95% CI 0.15–0.96). The adjusted odds of being anti-DENV seropositive was higher among anti-CHIKV seropositive adults, and vice versa (aOR 1.94, 95% CI 1.09–3.44), suggesting co-circulation of these viruses. DENV and CHIKV exposure is lower in Qatar and MENA nationals compared to Asian nationals suggesting a lower burden of DENV and CHIKV disease in the MENA. Antibodies to both viruses were detected in nationals from most MENA countries, supporting the need to better understand the regional epidemiology of these viruses.

Laser scanning optical investigations of dengue virus infection from antibody fluorescence of human blood

Dengue is the most important arthropod-borne viral disease of public health significance today, and the geographic distribution includes more than 100 countries worldwide. A definite diagnosis of dengue virus infection is based on clinical criteria supported by antibody tests of whole blood. In this study, we report on dengue infected whole blood diagnostics using laser scanning confocal microscopy (LSCM) over a laser excitation of 488, 543 and 633 nm wavelengths. Based on our findings, the system has potential applications in the detection and quantification of dengue virus infected cells, antigens and antibodies in blood in vitro. Analysis of about 100 infected patients shows that the primary organism (or pathogen), characteristic of dengue infection, causes a significant biochemical change in the nature of human blood in which it resides, and, due to which, fluorescence increases in dengue samples. Thus, the use of a reliable and sensitive rapid diagnostic test using LSCM can simultaneously detect the presence of anti-dengue IgG and IgM antibodies, which is a breakthrough in diagnosis and of great clinical use. The antibodies for dengue continue to be distinguishable for a couple of months after infection. This change in blood fluorescence may be a significant contribution from light scattering through antibodies produced against the dengue antigen.

Asymptomatic dengue virus cases in misiones, Argentina: a seroprevalence study in the university population

Dengue infection may be asymptomatic and may produce the typical symptoms of a benign illness or serious hemorrhagic and often fatal symptoms. Asymptomatic cases are statistically relevant and quite variable depending on the geographic area under study. However, there are no reports of asymptomatic population infected by the dengue virus in Misiones. In this study, 288 samples were analyzed, and the IgG anti dengue antibodies detected accounted for 6.6% of cases, while 89% corresponded to individuals who lived with people diagnosed or suspected of having contracted dengue, p= <0.001.

Can rapid dengue diagnostic kits be trusted? A comparative study of commercially available rapid kits for serodiagnosis of dengue fever.

BACKGROUND:Dengue virus infection is an important emerging disease of the tropical and subtropical regions and is mainly diagnosed by serological detection of NS1 antigen and IgM antidengue antibodies. Since enzyme-linked immunosorbent assay (ELISA) facilities are not easily available at most diagnostic centers, so most of them use various commercially available rapid diagnostic tests (RDTs) kits.AIMS AND OBJECTIVES:This study was designed to access the diagnostic accuracy of four commercially available and widely used RDTs for serodiagnosis of dengue virus infection in Indian laboratories.SUBJECTS AND METHODS:The study was conducted at Department of Microbiology, G.S.V.M Medical College, Kanpur, India, to estimate the sensitivity and specificity of following RDTs: (1) Dengue Cassette (Panbio, Australia), (2) Bioline Dengue Duo (SD Diagnostics, Korea), (3) Dengue Day 1 test (J Mitra and Co., India), and (4) Dengucheck Duo (Tulip Diagnostics, India) on 72 confirmed dengue serum samples that were positive by dengue reverse transcription-polymerase chain reaction, dengue NS1, and IgM ELISA along with 80 serum samples from nondengue febrile illness patients.RESULTS:The majority of the RDTs demonstrated low sensitivity but good specificity for detecting NS1 antigen. Detection of antidengue IgM antibodies by RDTs demonstrated low sensitivity ranging from 27.8% to 77.7%. However, specificity was generally higher (50%–86.2%) and more consistent across the assays.CONCLUSION:The study results differed markedly from the RDTs manufacturers' claimed performance characteristics. Therefore, the RDT results should be interpreted cautiously and ELISA should be performed as far as possible for serodiagnosis of dengue virus infection.

Selection and Characterization of Anti-Dengue NS1 Single Domain Antibodies

Reliable detection and diagnosis of dengue virus (DENV) is important for both patient care and epidemiological control. Starting with a llama immunized with a mixture of recombinant nonstructural protein 1 (NS1) antigen from the four DENV serotypes, a phage display immune library of single domain antibodies was constructed and binders selected which exhibited specificity and affinity for DENV NS1. Each of these single domain antibodies was evaluated for its binding affinity to NS1 from the four serotypes, and incorporated into a sandwich format for NS1 detection. An optimal pair was chosen that provided the best combination of sensitivity for all four DENV NS1 antigens spiked into 50% human serum while showing no cross reactivity to NS1 from Zika virus, yellow fever virus, tick-borne encephalitis virus, and minimal binding to NS1 from Japanese encephalitis virus and West Nile virus. These rugged and robust recombinant binding molecules offer attractive alternatives to conventional antibodies for implementation into immunoassays destined for resource limited locals.

Low frequency of asymptomatic dengue virus-infected donors in blood donor centers during the largest dengue outbreak in Taiwan.

To determine the prevalence of asymptomatic dengue virus-infected blood donors during the largest dengue outbreak in Taiwan history occurred in 2015, we examined the evidence of dengue virus (DENV) infection by the detection of DENV RNA genome using real-time reverse transcription-polymerase chain reaction (real-time RT-PCR), DENV NS1 antigen using rapid diagnosis test (RDT) and anti-dengue antibody using IgM/IgG capture enzyme-linked immunosorbent assay (capture ELISA) and RDT in eight thousand serum samples from blood donations to the blood centers of the Taiwan Blood Services Foundation (TBSF) in Kaohsiung City and Tainan City during the largest dengue outbreak in Taiwan history occurred in 2015. Only one serum sample was positive for DENV RNA detection by using dengue-specific real-time RT-PCR, the virus was DENV-2 determined by serotype-specific real-time RT-PCR and sequencing, and the DENVs in the serum were confirmed as being infectious by a plaque assay. The recipient of this blood did not develop any dengue fever symptom on follow-up. None of the samples was NS1 RDT-reactive. Seventeen IgM-positive samples were identified. There was a low prevalence of asymptomatic confirmed or probable DENV-infected blood donors in our study (0.013% and 0.21%, respectively), and no symptomatic transfusion-transmitted dengue (TT dengue) was developed during the largest dengue outbreak in Taiwan history in highly endemic areas and periods.

Seroprevalence of dengue fever and the associated sociodemographic, clinical, and environmental factors in Makkah, Madinah, Jeddah, and Jizan, Kingdom of Saudi Arabia

This study aimed to estimate the seroprevalence of anti-dengue IgG antibodies in Makkah, Al Madinah, Jeddah, and Jizan; and to identify the associated demographic, clinical, and environmental independent risk factors. A community-based household serosurvey conducted between September 20, 2016 and January 31, 2017. A multi-stage stratified cluster sampling was used to select 6596 participants from Makkah, Madinah, Jeddah, and Jizan. Blood samples were drawn from all participants to detect anti-dengue IgG antibodies. A semi-structured questionnaire was used to collect information on demographic, clinical, and environmental data. Multivariate logistic regression was carried out to identify independent risk factors of dengue seropositivity. The dengue seroprevalence (95% confidence intervalI) was 26.7% (25.6%, 27.8%), with the highest (33.6%) and lowest (14.8%) rates in Jizan and Madinah, respectively, and reaching 50% or more in several districts of the four cities. Demographic predictors of seroprevalence included: dwelling in Makkah (odds ratio [OR] = 2.19, p < 0.001) or Jizan (OR = 2.17, p < 0.001); older age (OR = 3.91, p < 0.001 for age>30 years); housing type (OR = 1.84 and 1.82, p < 0.001 for popular and social houses, respectively); and number of household occupants (OR = 0.86 and 0.71 for 6–10 [p = 0.042] and 11–20 [p = 0.002] occupants, respectively). Environmental predictors included the absence of pest control works in residency area (OR = 1.39, p = 0.002), presence of mosquitoes in the home (OR = 1.39, p = 0.001), and absence of awareness campaigns (OR = 1.97, p < 0.001). One in four inhabitants of the Western region of Saudi Arabia was seropositive for the dengue virus. Implementation of behavior-based educational programs is recommended, involving the population in the identification and eradication of vector sources and promoting appropriate behaviors that prevent the spread.