In Vitro Cleavage Assay to Characterize DENV NS2B3 Antagonism of cGAS.

cGAS is a key cytosolic dsDNA receptor that senses viral infection and elicits interferon production through the cGAS-cGAMP-STING axis. cGAS is activated by dsDNA from viral and bacterial origins as well as dsDNA leaked from damaged mitochondria and nucleus. Eventually, cGAS activation launches the cell into an antiviral state to restrict the replication of both DNA and RNA viruses. Throughout the long co-evolution, viruses devise many strategies to evade cGAS detection or suppress cGAS activation. We recently reported that the Dengue virus protease NS2B3 proteolytically cleaves human cGAS in its N-terminal region, effectively reducing cGAS binding to DNA and consequent production of the second messenger cGAMP. Several other RNA viruses likely adopt the cleavage strategy. Here, we describe a protocol for the purification of recombinant human cGAS and Dengue NS2B3 protease, as well as the in vitro cleavage assay.

Beyond the Bite: Detailed findings on Chikungunya and Dengue co-detection in Punjab, North India - clinical insights and diagnostic challenges.

OBJECTIVES: The co-circulation of Chikungunya virus (CHIKV) and Dengue virus (DENV) in India poses a challenge for the diagnosing clinician, as they share similar clinical signs and symptoms and geographical distribution. Both arthropod-borne viruses are maintained in the environment by the Aedes mosquito, commonly found in tropical countries including India. Here we aim to investigate the clinical and laboratory aspects of Chikungunya/Dengue suspected cases in Punjab, India during 2021-2022, focusing on the differential diagnosis of Dengue. METHODS: All suspected cases were submitted to serological differential diagnosis approaches to arboviruses like Chikungunya and Dengue. For the detection of Chikungunya Infection, CHIK IgM Capture ELISA was employed. Whereas, for Dengue NS1 antigen ELISA and IgM Capture ELISA assays were employed. RESULTS: A total of 370 cases suspected of arboviral infection were investigated and 38.3% (142/370) were confirmed as Chikungunya. Chikungunya cases were slightly more prevalent in males (54%) and the most frequently affected age group was adults between 16 and 30 years old (45.7%). Polyarthralgia affected 79.5% of patients, 63.3% exhibited headache and 50% presented with retro-orbital pain. 28.9% (107/370) had serological evidence of DENV exposure by detection of specific anti-DENV IgM or NS1 and 9.1% (34/370) cases of co-detection of Chikungunya and Dengue were reported. Urban populations had a higher infection rate of co-detection of Chikungunya and Dengue than rural populations with 83% versus 17%, respectively. CONCLUSIONS: Despite an initial clinical diagnosis of Dengue, most patients with fever and arthralgia were serologically confirmed as Chikungunya cases, with a notable prevalence of CHIKV/DENV co-detection. Strengthening differential diagnosis of circulating arboviruses is crucial for improving patient care and enhancing vector control and environmental management strategies.

Understanding the unprecedented 2023 dengue outbreak in Bangladesh: a data-driven analysis.

Objectives: This study aims to elucidate the epidemiological characteristics, spatial distribution, and potential contributing factors associated with the 2022-2023 dengue outbreak in Bangladesh. Methods: We retrospectively analyzed dengue fever cases reported by national health surveillance systems, focusing on incidence, geographical spread, and fatalities. Statistical methods were used to explore correlations between population density, healthcare capacity, and disease prevalence. Results: Our study revealed that in 2023, dengue cases and deaths surged five-fold (from 62,382 to 320,835) and nearly six-fold (from 281 to 1699) compared with 2022. Major cities such as Dhaka and Chittagong emerged as epicenters with significantly higher caseloads and mortality rates. The analysis identified a strong positive correlation between population density and disease prevalence, suggesting urbanization as a contributing factor. In addition, a shift in the peak dengue season from August to September was observed. Furthermore, disparities in health care infrastructure were identified, with densely populated areas experiencing critical shortages of hospital beds, potentially impacting fatality rates. Conclusions: This unprecedented dengue outbreak in Bangladesh highlights the need for a multifaceted approach. Prioritizing vector control, targeted public awareness in identified hotspots, addressing healthcare resource inequities, and further research on environmental and demographic determinants of transmission are crucial for mitigating future outbreaks in Bangladesh.

Dengue virus and the 2024 Paris Olympics.

The 2024 Paris Olympics and Paralympics face concerns over dengue virus transmission, despite Paris's lower mosquito activity. Preventive measures include eliminating breeding sites, insecticide spraying, and public awareness. Health systems will monitor and respond to cases. Large gatherings like the Olympics can amplify disease spread, as seen with Zika in Rio 2016. Recent reports confirm dengue presence in Europe, highlighting global risks. While Paris's overall dengue risk is low, even a few cases could impact global health. Collaboration among health authorities, researchers, and event organizers is crucial to ensure participant and public safety during the games.

Estimating dengue disease and economic burden to inform municipal-level policymakers: Method for a pragmatic city-level observational cohort study.

Background: Recent trials have confirmed the effectiveness of promising dengue control technologies - two vaccines and Wolbachia. These would generally be applied at the municipal level. To help local officials decide which, if any, control strategy to implement, they need affordable, timely, and accurate data on dengue burden. Building on our previous work in Mexico, Indonesia, and Thailand, we developed a streamlined prospective method to estimate dengue burden at the municipal level quickly, accurately, and efficiently. Methods: The method entails enrolling and repeatedly interviewing 100 patients with laboratory-confirmed dengue. They will be selected after screening and testing about 1,000 patients with clinical dengue. The method will capture both acute and chronic effects relating to disease, economic burden, and psychological impacts (presenteeism). The total time requirements are 1.5 years, comprised of 0.25 years for planning and approvals, 1 year for data collection (a full dengue cycle), and 0 .25 years for data cleaning and analysis. A collaboration with municipal and academic colleagues in the city of Semarang, Central Java, Indonesia shows how the method could be readily applied in Indonesia's eighth largest city (population 1.8 million). Conclusions: Many surveillance studies gather only information on numbers of cases. This proposed method will provide a comprehensive picture of the dengue burden to the health system, payers, and households at the local level.

Telas de proteção em habitações para prevenção de doenças transmitidas por mosquitos: sinopse baseada em evidências

Contexto e objetivo: A transmissão de doenças por mosquitos afeta a população e a economia de todo o mundo. Há um número considerável de doenças que podem ser transmitidas por mosquitos, com destaque para a malária e a dengue, endêmica em regiões tropicais. Evidentemente, medidas preventivas são imprescindíveis para a redução da transmissão. Avaliar as evidências de efetividade das telas de proteção com e sem inseticida para prevenção de doenças transmitidas por mosquitos. Métodos: Trata-se de sinopse baseada em evidências. Procedeu-se à busca por estudos que associavam o uso de telas de proteção contra mosquitos à redução do contágio de doenças transmitidas por mosquitos em três bases de dados: PubMed (1966-2024), Portal BVS (1982-2024) e Epistemonikos (2024) e também no metabuscador de evidências TRIP DATABASE (2024). O desfecho de análise envolveu a efetividade das telas de proteção na redução de doenças transmitidas por mosquitos. Resultados: Foram encontradas 307 citações. Seis estudos (1 revisão sistemática e 5 ensaios clínicos) foram incluídos. Discussão: A maioria dos estudos envolveu a colocação de telas de proteção com inseticida, havendo evidência de alta certeza para redução de mortalidade por malária e redução na entrada de mosquitos nas habitações, mesmo com redes sem inseticida. Conclusões: Embora não haja robustez na evidência da efetividade das telas de proteção sem inseticidas contra mosquitos transmissores de doenças, o que demanda a necessidade de realização de novos estudos prospectivos, parece lícita e benéfica a utilização de telas de proteção em regiões endêmicas para doenças transmitidas por esses vetores.

Knowledge, attitude, and practice towards dengue fever among medical students in Sudan: A cross-sectional study.

Background: Dengue fever, a neglected tropical disease, exhibits a total seroprevalence of 27% in Sudan. During the recent outbreak, medical students actively participated in health education and community outreach programs. This study aims to assess the knowledge, attitude, and practice (KAP) of medical students regarding Dengue Fever (DF) during its current outbreak in Sudan. Design and methods: We conducted a cross-sectional study involving medical students from more than 15 Sudanese universities, encompassing 11 states. Data collection took place between October 23 and November 10, 2023, utilizing a Google Form questionnaire. A convenient sampling technique was employed to reach students due to its accessibility and feasibility during the ongoing conflict in Sudan. Correlation and regression analyses were used to identify predictors of KAP and establish associations between KAP domains and socio-demographic factors. Results: Of 694 medical students, 11% reported a previous infection with DF. Among the participants, 58% demonstrated good knowledge, 74.1% exhibited a positive attitude, and less than half (46.7%) demonstrated good preventive practices. There was a positive correlation between knowledge and attitude levels with the practice level (p < 0.001). Gender, age, and previous DF infection emerged as significant predictors (p < 0.05) of the overall KAP level in multiple logistic regression. Conclusions: Our study revealed a commendable level of knowledge and attitude but a suboptimal level of practice regarding DF among medical students. This inadequacy in practice, particularly among medical students who are expected to set an example, is concerning and warrants further investigation.

The global incidence and trends of three common flavivirus infections (Dengue, yellow fever, and Zika) from 2011 to 2021.

Background: Flavivirus pose a continued threat to global health, yet their worldwide burden and trends remain poorly quantified. We aimed to evaluate the global, regional, and national incidence of three common flavivirus infections (Dengue, yellow fever, and Zika) from 2011 to 2021. Methods: Data on the number and rate of incidence for the three common flavivirus infection in 204 countries and territories were retrieved from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021. The estimated annual percent change (EAPC) was calculated to quantify the temporal trend during 2011-2016, 2016-2019, and 2019-2021, respectively. Results: In 2021, an estimated 59,220,428 individuals were infected globally, comprising 58,964,185 cases of dengue, 86,509 cases of yellow fever, and 169,734 cases of Zika virus infection. The age-standardized incidence rate (ASIR) of the three common flavivirus infections increased by an annual average of 5.08% (95% CI 4.12 to 6.05) globally from 2011 to 2016, whereas decreased by an annual average of -8.37% (95% CI -12.46 to -4.08) per year between 2016 to 2019. The ASIR remained stable during 2019-2021, with an average change of 0.69% (95% CI -0.96 to 2.37) per year globally for the three common flavivirus infections. Regionally, the burden of the three common flavivirus infections was primarily concentrated in those regions with middle income, such as South Asia, Southeast Asia, and Tropical Latin America. Additionally, at the country level, there was an inverted "U" relationship between the SDI level and the ASI. Notably, an increase in the average age of infected cases has been observed worldwide, particularly in higher-income regions. Conclusion: Flavivirus infections are an expanding public health concern worldwide, with considerable regional and demographic variation in the incidence. Policymakers and healthcare providers must stay vigilant regarding the impact of COVID-19 and other environmental factors on the risk of flavivirus infection and be prepared for potential future outbreaks.

Prevalence of Dengue, Zika, and Chikungunya Virus Infections among Mosquitoes in Asia: A Systematic Review and Meta-Analysis.

BACKGROUND: Dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) continue to pose significant public health risks. This study aims to assess the prevalence of these arbovirus infections in field-caught mosquitoes across Asia. METHODS: Studies published after the year 2000 on DENV, ZIKV, and/or CHIKV infections in Asian mosquitoes were identified from Embase, Scopus, PubMed, and Ovid. A random-effects model estimated the pooled prevalence, defined as the overall prevalence from included studies, adjusted for variability among the studies. Meta-regression models were used to evaluate the association between predictors and their prevalence. RESULTS: A total of 2,529 articles were retrieved; 57 met the inclusion criteria. Pooled prevalence of DENV, ZIKV, and CHIKV infections in Asian mosquitoes were 5.85%, 2.15%, and 1.26%, respectively. Subgroup analysis revealed varying DENV prevalence across regions: East Asia (3.32%), South Asia (5.26%), and Southeast Asia (6.92%). Univariate regression analysis demonstrated significant associations between mosquito capture site and DENV prevalence (p<0.001), and between study region and ZIKV prevalence (p=0.005). However, no significant predictors were identified for CHIKV prevalence. CONCLUSION: Our findings provide reference pooled summary estimates of arbovirus infections in mosquitoes, offering crucial insight into the regional disease burden and guiding the development and implementation of arbovirus surveillance in mosquitoes.

Comparison of the sensitivity and specificity of commercial anti-dengue virus IgG tests to identify persons eligible for dengue vaccination.

The Advisory Committee on Immunization Practices (ACIP) recommended that dengue pre-vaccination screening tests for Dengvaxia administration have at least 98% specificity and 75% sensitivity. This study evaluates the performance of commercial anti-DENV IgG tests to identify tests that could be used for pre-vaccination screening. First, for seven tests, we evaluated sensitivity and specificity in early convalescent dengue virus (DENV) infection, using 44 samples collected 7-30 days after symptom onset and confirmed by RT-PCR. Next, for the five best-performing tests and two additional tests (with and without an external test reader) that became available later, we evaluated performance to detect past dengue infection among a panel of 44 specimens collected in 2018-2019 from healthy 9- to 16-year-old children from Puerto Rico. Finally, a full-scale evaluation was done with the four best-performing tests using 400 specimens from the same population. We used virus focus reduction neutralization test and an in-house DENV IgG ELISA as reference standards. Of seven tests, five showed ≥75% sensitivity in detecting anti-DENV IgG in early convalescent specimens with low cross-reactivity to the Zika virus. For the detection of previous DENV infections, the tests with the highest performance were the Euroimmun NS1 IgG ELISA (sensitivity 84.5%, specificity 97.1%) and CTK Dengue IgG rapid test R0065C with the test reader (sensitivity 76.2% specificity 98.1%). There are IgG tests available that can be used to accurately classify individuals with previous DENV infection as eligible for dengue vaccination to support safe vaccine implementation. IMPORTANCE: The Advisory Committee on Immunization Practices (ACIP) has set forth recommendations that dengue pre-vaccination screening tests must exhibit at least 98% specificity and 75% sensitivity. Our research rigorously assesses the performance of various commercial tests against these benchmarks using well-characterized specimens from Puerto Rico. The findings from our study are particularly relevant given FDA approval and ACIP recommendation of Sanofi Pasteur's Dengvaxia vaccine, highlighting the need for accurate pre-vaccination screening tools.

Intra-Phenotypic and -Genotypic Variations of Beauveria bassiana (Bals.) Vuill. Strains Infecting Aedes aegypti L. Adults.

Beauveria bassiana has potential for Aedes aegypti biological control. However, its efficacy depends on the strain's geographic location, host susceptibility, and virulence. The present study aimed to evaluate the effectiveness of B. bassiana strain BBPTG4 conidia in controlling Ae. aegypti adults and its detection via introns profile on exposed mosquito corpses. Morphologic characteristics among strains were highly similar. Comprehensive testing of these strains demonstrated that BBPT4 exhibited the ideal biological activity for Ae. aegypti control, with a median lethal time (TL50) of 7.5 d compared to ~3 d and ~10 d for BB01 and BB37 strains, respectively. Infected mosquitoes died after GHA and BBPTG4 exposure, and corpses were analyzed for infecting strains detection. Differences among the seven evaluated strains were determined, assessing five different insertion group I intron profiles in BBTG4, BB01, GHA, BB37, and BB02 strains. Mosquitoes infected by BBPTG4 and non-exposed (negative control) intron profiles were obtained. We detected the presence of introns in the BBPTG4 strain, which were not present in non-exposed mosquitoes. In conclusion, B. bassiana strains showed similarities in terms of their cultural and microscopic morphological characteristics and biologicals virulence level, but different intron profiles. BBPTG4 strain-infected Ae. aegypti adult corpses, showing specific amplicons, enabled us to identify B. bassiana at the strain level among infected mosquitoes. However, monitoring and detection of field-infected insects is essential for further verification.

Aedes aegypti Mosquito Probing Enhances Dengue Virus Infection of Resident Myeloid Cells in Human Skin.

The most prevalent arthropod-borne viruses, including the dengue viruses, are primarily transmitted by infected mosquitoes. However, the dynamics of dengue virus (DENV) infection and dissemination in human skin following Aedes aegypti probing remain poorly understood. We exposed human skin explants to adult female Ae. aegypti mosquitoes following their infection with DENV-2 by intrathoracic injection. Skin explants inoculated with a similar quantity of DENV-2 by a bifurcated needle were used as controls. Quantitative in situ imaging revealed that DENV replication was greatest in keratinocytes in the base of the epidermis, accounting for 50-60% of all infected cells regardless of the route of inoculation. However, DENV inoculation by Ae. aegypti probing resulted in an earlier and increased viral replication in the dermis, infecting twice as many cells at 24 h when compared to needle inoculation. Within the dermis, enhanced replication of DENV by Ae. aegypti infected mosquitoes was mediated by increased local recruitment of skin-resident macrophages, dermal dendritic cells, and epidermal Langerhans cells relative to needle inoculation. An enhanced but less pronounced influx of resident myeloid cells to the site of mosquito probing was also observed in the absence of infection. Ae. aegypti probing also increased recruitment and infection of dermal mast cells. Our findings reveal for the first time that keratinocytes are the primary targets of DENV infection following Ae. aegypti inoculation, even though most of the virus is inoculated into the dermis during probing. The data also show that mosquito probing promotes the local recruitment and infection of skin-resident myeloid cells in the absence of an intact vasculature, indicating that influx of blood-derived neutrophils is not an essential requirement for DENV spread within and out of skin.

Detection of Dengue Virus 1 and Mammalian Orthoreovirus 3, with Novel Reassortments, in a South African Family Returning from Thailand, 2017.

In July 2017, a family of three members, a 46-year-old male, a 45-year-old female and their 8-year-old daughter, returned to South Africa from Thailand. They presented symptoms consistent with mosquito-borne diseases, including fever, headache, severe body aches and nausea. Mosquito bites in all family members suggested recent exposure to arthropod-borne viruses. Dengue virus 1 (Genus Orthoflavivirus) was isolated (isolate no. SA397) from the serum of the 45-year-old female via intracerebral injection in neonatal mice and subsequent passage in VeroE6 cells. Phylogenetic analysis of this strain indicated close genetic identity with cosmopolitan genotype 1 DENV1 strains from Southeast Asia, assigned to major lineage K, minor lineage 1 (DENV1I_K.1), such as GZ8H (99.92%) collected in November 2018 from China, and DV1I-TM19-74 isolate (99.72%) identified in Bangkok, Thailand, in 2019. Serum samples from the 46-year-old male yielded a virus isolate that could not be confirmed as DENV1, prompting unbiased metagenomic sequencing for virus identification and characterization. Illumina sequencing identified multiple segments of a mammalian orthoreovirus (MRV), designated as Human/SA395/SA/2017. Genomic and phylogenetic analyses classified Human/SA395/SA/2017 as MRV-3 and assigned a tentative genotype, MRV-3d, based on the S1 segment. Genomic analyses suggested that Human/SA395/SA/2017 may have originated from reassortments of segments among swine, bat, and human MRVs. The closest identity of the viral attachment protein σ1 (S1) was related to a human isolate identified from Tahiti, French Polynesia, in 1960. This indicates ongoing circulation and co-circulation of Southeast Asian and Polynesian strains, but detailed knowledge is hampered by the limited availability of genomic surveillance. This case represents the rare concurrent detection of two distinct viruses with different transmission routes in the same family with similar clinical presentations. It highlights the complexity of diagnosing diseases with similar sequelae in travelers returning from tropical areas.

Infection Dynamics of Dengue Virus in Caco-2 Cells Depending on Its Differentiation Status.

Dengue virus (DENV), from the Flaviviridae family, is the causative agent of dengue fever and poses a significant global health challenge. The virus primarily affects the vascular system and liver; however, a growing body of evidence suggests its involvement in the gastrointestinal (GI) tract, contributing to clinical symptoms such as abdominal pain, vomiting, and diarrhea. However, the mechanisms underlying DENV infection in the digestive system remain largely unexplored. Prior research has detected viral RNA in the GI tissue of infected animals; however, whether the dengue virus can directly infect human enterocytes remains unclear. In this study, we examine the infectivity of human intestinal cell lines to the dengue virus and their subsequent response. We report that the Caco-2 cell line, a model of human enterocytes, is susceptible to infection and capable of producing viruses. Notably, differentiated Caco-2 cells exhibited a lower infection rate yet a higher level of virus production than their undifferentiated counterparts. These findings suggest that human intestinal cells are a viable target for the dengue virus, potentially elucidating the GI symptoms observed in dengue fever and offering a new perspective on the pathogenetic mechanisms of the virus.

New biomarkers for liver involvement by dengue infection in adult Vietnamese patients: a case-control study.

Liver injury with marked elevation of aspartate aminotransferase enzyme (AST) is commonly observed in dengue infection. To understand the pathogenesis of this liver damage, we compared the plasma levels of hepatic specific, centrilobular predominant enzymes (glutamate dehydrogenase, GLDH; glutathione S transferase-α, αGST), periportal enriched 4-hydroxyphenylpyruvate dioxygenase (HPPD), periportal predominant arginase-1 (ARG-1), and other non-specific biomarkers (paraoxonase-1, PON-1) in patients with different outcomes of dengue infection. This hospital-based study enrolled 87 adult dengue patients, stratified into three groups based on plasma AST levels (< 80, 80-400, > 400 U/L) in a 1:1:1 ratio (n = 40, n = 40, n = 40, respectively. The new liver enzymes in the blood samples from the 4th to 6th days of their illness were measured by commercial enzyme-linked immunosorbent assay (ELISA) or colorimetric kits. Based on the diagnosis at discharge days, our patients were classified as 40 (46%) dengue without warning signs (D), 35 (40.2%) dengue with warning signs (DWS), and 11 (12.6%) severe dengue (SD) with either shock (two patients) or AST level over 1000 U/L (nine patients), using the 2009 WHO classification. The group of high AST (> 400 U/L) also had higher ALT, GLDH, ARG-1, and HPPD than the other groups, while the high (> 400 U/L) and moderate (80-400 U/L) AST groups had higher ALT, αGST, ARG-1, and HPPD than the low AST group (< 80 U/L). There was a good correlation between AST, alanine aminotransferase enzyme (ALT), and the new liver biomarkers such as GLDH, αGST, ARG-1, and HPPD. Our findings suggest that dengue-induced liver damage initiates predominantly in the centrilobular area toward the portal area during the dengue progression. Moreover, these new biomarkers should be investigated further to explain the pathogenesis of dengue and to validate their prognostic utility.

Dynamics and Efficacy: A Comprehensive Evaluation of the Advanced Dengue Fever Surveillance and Early Warning System in Ningbo City, 2023.

Objective: To conduct a comprehensive evaluation of the Dengue Fever Surveillance and Early Warning System deployed in Ningbo City during 2023, focusing on its capacity for timely identification and reporting of dengue fever cases, particularly imported cases from endemic regions. Methods: A detailed data of patient clinical features and blood profile trends was collected from clinical records and surveillance reports, focusing on the rapid diagnostic processes and surveillance rigor. This study assessed the effectiveness of the system in identifying and reporting dengue cases and identified the limitations of the existing framework through a basic statistical approach. Results: The system demonstrated timely identification and reporting of dengue fever cases, with a particular emphasis on imported cases. However, several limitations were identified, including the need for more precise monitoring criteria and improved coordination with medical entities. Conclusion: The study underscores the critical role of public health bodies in managing disease outbreaks and advocates for enhanced methodologies to refine epidemic control efforts. The findings contribute to the advancement of early warning mechanisms and the improvement of proactive infectious disease monitoring in metropolitan environments, providing valuable insights for fortifying the Dengue Fever Surveillance and Early Warning System in Ningbo City.

Commercial Enzyme-Linked Immunosorbent Assay Kit Is Useful for Detection of Recombinant and Secretory Nonstructural-1 Protein Antigen of Dengue Virus.

Background: Dengue is a mosquito-borne tropical disease, caused by the Dengue virus (DENV). It has become a severe problem and is a rising threat to public health. In this study, we have evaluated commercial Merilisa i Dengue NS1 Antigen kit (Meril LifeSciences India Pvt. Ltd.) to detect recombinant dengue virus 2 NS1 antigen (rDNS1Ag) and secreted forms of NS1 antigen (sDNS1Ag). Methods: To determine the detection limit of the kit, 100 nanogram (ng) to 0.001 ng rDNS1Ag was tested. The sensitivity and specificity of the kit was determined using recombinant NS1 antigens of all serotypes of DENV and other flaviviruses. For testing sDNS1Ag, the culture supernatant of the Vero cell lines infected with DENV-2 was tested. Further, a spiking experiment was carried out to check the sensitivity of the kit to detect rDNS1Ag in the pools of Aedes aegypti mosquitoes. Results: It was observed that the kit can detect the rDNS1Ag at 1 ng concentration. The kit was sensitive to detect NS1 antigen of DENV-1, DENV-2 and DENV-3 serotypes and specific for detection of only DNS1Ag as it did not cross-react with NS1 antigen of flaviviruses. The kit was sensitive to detect rDNS1Ag in the mosquito pools as well. In addition, the kit was able to detect the sDNS1Ag in Vero cell culture supernatant. Conclusions: Overall, we observed that the Merilisa i Dengue NS1 Ag kit is sensitive and specific for the detection of DNS1Ag both in recombinant and secretory forms.

Post-dengue mucormycosis- a novel entity or COVID-19 associated in disguise?

Dengue is a disease endemic to tropical countries such as India. In the past two years, cases of COVID-19 associated Mucormycosis have become a commonly encountered phenomenon. However, cases of dengue associated Mucormycosis have not found a significant mention in the literature yet. We, therefore, report two such cases in which Mucormycosis developed after recovery from Dengue fever and try to dig into the possible mechanism behind such an occurrence.

Differential Proteomic Profiling at Different Phases of Dengue Infection: An Intricate Insight from Proteins to Pathogenesis.

Dengue fever is a rapidly emerging tropical disease and an important cause of morbidity in its severe form worldwide. A wide spectrum of the pathophysiology is associated with the transition of dengue fever to severe dengue, which is driven by the host immune response and might reflect in patients' proteome profile. This study aims to analyze the plasma from different phases of dengue-infected patients at two time points. A mass-spectrometry-based proteomic approach was utilized to understand the involvement of probable candidate proteins toward developing a more severe, hemorrhagic form of dengue fever. Dengue-infected hospital-admitted patients with <5 days of fever were included in this study. Patient samples from the acute phase were screened for the presence of NS1 antigen using ELISA and subjected to molecular serotyping. Dengue molecular serotype-confirmed patient samples, pairwise from acute and critical phases with healthy control were subjected to qualitative and quantitative proteomic analysis, and then pathway analysis was performed. The protein-protein interaction network between the dengue virus and host proteins was depicted in the search for proteins associated with severe dengue pathophysiology. An array of apolipoprotein, cytokines, and endothelial proteins in association with virus replication and endothelial dysfunction were validated as biomolecules involved in severe dengue pathophysiology.