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1.
Viruses ; 15(6)2023 06 20.
Article in English | MEDLINE | ID: mdl-37376696

ABSTRACT

Human enterovirus causes various clinical manifestations in the form of rashes, febrile illness, flu-like illness, uveitis, hand-foot-mouth disease (HFMD), herpangina, meningitis, and encephalitis. Enterovirus A71 and coxsackievirus are significant causes of epidemic HFMD worldwide, especially in children aged from birth to five years old. The enterovirus genotype variants causing HFMD epidemics have been reported increasingly worldwide in the last decade. We aim to use simple and robust molecular tools to investigate human enteroviruses circulating among kindergarten students at genotype and subgenotype levels. With the partial 5'-UTR sequencing analysis as a low-resolution preliminary grouping tool, ten enterovirus A71 (EV-A71) and coxsackievirus clusters were identified among 18 symptomatic cases and 14 asymptomatic cases in five kindergartens in Bangkok, Thailand, between July 2019 and January 2020. Two occurrences of a single clone causing an infection cluster were identified (EV-A71 C1-like subgenotype and coxsackievirus A6). Random amplification-based sequencing using MinION (Oxford Nanopore Technology) helped identify viral transmission between two closely related clones. Diverse genotypes co-circulating among children in kindergartens are reservoirs for new genotype variants emerging, which might be more virulent or better at immune escape. Surveillance of highly contagious enterovirus in communities is essential for disease notifications and controls.


Subject(s)
Enterovirus A, Human , Enterovirus Infections , Enterovirus , Hand, Foot and Mouth Disease , Child , Humans , Thailand/epidemiology , Enterovirus/genetics , Hand, Foot and Mouth Disease/epidemiology , Genotype , China/epidemiology
2.
Trop Med Infect Dis ; 8(5)2023 May 17.
Article in English | MEDLINE | ID: mdl-37235332

ABSTRACT

The frequency of Zika virus (ZIKV)-specific IgA and IgM and the cytokine expression profile of ZIKV-infected patients in hyperendemic areas remain unclear. This study investigated the rates of ZIKV non-structural protein 1 (NS1)-specific IgA and IgM and evaluated serum cytokine levels of ZIKV and Dengue virus (DENV) cases in Thailand to identify potential diagnostic biomarkers, elucidate the immunity against ZIKV and DENV, and investigate the association between cytokine levels and ZIKV symptoms. Low rates of positivity for ZIKV NS1-specific IgA and IgM were detected in our study. ZIKV NS1 IgA/M (11%, 11/101) in combination was more frequently detected than ZIKV NS1 IgM (2%, 2/101) or ZIKV NS1 IgA (4%, 4/96) alone, especially in acute ZIKV cases with previous DENV exposure (14%, 10/72). Cytokine analysis showed that both ZIKV and DENV infections induced polyfunctional immunity, and the latter triggered more prolonged responses. The existence of significant differences in IL-4 and IL-10 levels between acute ZIKV and acute DENV cases suggested that IL-4 (p = 0.0176) and IL-10 (p = 0.0003) may represent biomarkers for acute ZIKV and acute DENV infections, respectively. Analysis of the association between increased cytokine levels and ZIKV symptoms indicated that CXCL10 (p = 0.0029) was associated with exanthema, while IL-5 (p = 0.0496) was linked to headache. The detection of ZIKV NS1 IgA and IgM in combination may enhance the diagnosis of early ZIKV infection, particularly when levels of IgM or IgA alone are low or undetectable. IL-4 and IL-10 may serve as targets for the development of diagnostic tools to detect ZIKV and DENV infections early, respectively, in flavivirus-endemic regions.

3.
Biomedicines ; 11(1)2023 Jan 16.
Article in English | MEDLINE | ID: mdl-36672734

ABSTRACT

Due to the lack of an effective therapeutic treatment to flavivirus, dengue virus (DENV) nonstructural protein 1 (NS1) has been considered to develop a vaccine owing to its lack of a role in antibody-dependent enhancement (ADE). However, both NS1 and its antibody have shown cross-reactivity to host molecules and have stimulated anti-DENV NS1 antibody-mediated endothelial damage and platelet dysfunction. To overcome the pathogenic events and reactogenicity, human monoclonal antibodies (HuMAbs) against DENV NS1 were generated from DENV-infected patients. Herein, the four DENV NS1-specific HuMAbs revealed the therapeutic effects in viral neutralization, reduction of viral replication, and enhancement of cell cytolysis of DENV and zika virus (ZIKV) via complement pathway. Furthermore, we demonstrate that DENV and ZIKV NS1 trigger endothelial dysfunction, leading to vascular permeability in vitro. Nevertheless, the pathogenic effects from NS1 were impeded by 2 HuMAbs (D25-4D4C3 and D25-2B11E7) and also protected the massive cytokines stimulation (interleukin [IL-]-1b, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-8, IL-9, IL-13, IL-17, eotaxin, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, Inducible protein-10, monocyte chemoattractant protein-1, macrophage inflammatory protein [MIP]-1 α, MIP-1ß, tumor necrosis factor-α, platelet-derived growth factor, and RANTES). Collectively, our findings suggest that the novel protective NS1 monoclonal antibodies generated from humans has multiple therapeutic benefits against DENV and ZIKV infections.

4.
Pathog Glob Health ; 117(1): 76-84, 2023 02.
Article in English | MEDLINE | ID: mdl-35708203

ABSTRACT

Circulating hsa-miRNA-126 (CmiR-126) has been reported to involve in the pathogenesis of many infectious diseases including dengue virus infection. However, no prior study has been conducted to describe more details in dengue-infected pediatric patients. This study aimed to describe CmiR-126-3p in dengue-infected pediatric patients during the febrile and convalescent phases. Additionally, the correlations between CmiR-126-3p and other relevant clinical laboratory factors were investigated. Sixty paired-serum specimens collected during febrile and convalescent phases were retrieved from patients with dengue fever (DF) (n = 30) and dengue hemorrhagic fever (DHF) (n = 30). Thirty paired-serum specimens collected from non-dengue acute febrile illness patients (AFI) were included as the control group. CmiR-126-3p was determined using reverse transcription quantitative real-time polymerase-chain reaction (RT-qPCR). Relative miRNA expression was calculated as 2-ΔCt using CmiR-16-5p for data normalization. CmiR-126-3p expression during febrile and convalescent phases in dengue-infected patients was significantly lower than AFI (p < 0.05). However, miRNA levels were not different (p > 0.05) compared between DF and DHF and between primary and secondary infection. CmiR-126-3p levels in DF in the convalescent were significantly higher than in the febrile phase (p = 0.025). No association between CmiR-126-3p and hematocrit, WBC level, platelet count, WBC differential count or dengue viral load was observed (p > 0.05). The data suggest that hsa-miR-126-3p involved in pathogenesis of dengue infection and may be a promising early and late biomarker for DENV infection. However, hsa-miR-126-3p alone cannot be used as a predictor for dengue severity.


Subject(s)
Circulating MicroRNA , MicroRNAs , Humans , Child , Southeast Asian People , MicroRNAs/genetics , Biomarkers , Real-Time Polymerase Chain Reaction
5.
Trop Med Infect Dis ; 7(11)2022 Nov 15.
Article in English | MEDLINE | ID: mdl-36422929

ABSTRACT

Arboviruses, particularly dengue virus (DENV), Zika virus (ZIKV), and Chikungunya virus (CHIKV), pose a growing threat to global public health. For disease burden estimation and disease control, seroprevalence studies are paramount. This study was performed to determine the prevalence of DENV, ZIKV, and CHIKV on healthy individuals aged from 1-55 years old in Bangphae district, Ratchaburi province, Thailand. Enzyme-linked immunosorbent assays (ELISAs) and rapid diagnostic tests (RDTs) were performed on archived samples from a dengue serological survey conducted from 2012-2015. All 2012 samples had been previously tested using an anti-DENV immunoglobulin (Ig)G ELISA, and 400 randomly selected samples stratified by age, sex, and residential area were assessed by an in-house anti-ZIKV IgG ELISA and a commercial anti-CHIKV IgG ELISA to determine virus-specific antibody levels. An RDT (Chembio DPP® ZCD IgM/IgG System) was also used to investigate the presence of antibodies against DENV, ZIKV, or CHIKV. The ELISA results indicate that the seroprevalences of DENV, ZIKV, and CHIKV were 84.3%, 58.0%, and 22.5%, respectively. The youngest age group had the lowest seroprevalence for all three arboviruses, and the seroprevalences for these viruses were progressively higher with increasing participant age. The DPP® IgG sensitivities, as compared with ELISAs, for DENV, ZIKV, and CHIKV were relatively low, only 43.92%, 25.86%, and 37.78%, respectively. The ELISA results indicate that 16% of the study population was seropositive for all three viruses. DENV had the highest seroprevalence. ZIKV and CHIKV were also circulating in Bangphae district, Ratchaburi province, Thailand. The DPP® ZCD rapid test is not sensitive enough for use in seroprevalence studies.

6.
Trop Med Infect Dis ; 7(10)2022 Sep 21.
Article in English | MEDLINE | ID: mdl-36287995

ABSTRACT

Zika virus (ZIKV) is a mosquito-borne flavivirus that has recently emerged as a global health threat. The rise in ZIKV infections has driven an increased incidence of neonates born with microcephaly or other neurological malformations. Therefore, screening for ZIKV infection can considerably impact pregnant women, especially during the first trimester. The majority of ZIKV infections are mild or asymptomatic, and clinical diagnosis is inaccurate. Moreover, given the high level of cross-reactivity among flaviviruses, serological approaches to distinguish ZIKV from dengue virus (DENV) infections are complicated. We used the combination of DENV and ZIKV nonstructural protein 1 (NS1) IgG enzyme-linked immunosorbent assay (ELISA) and ZIKV NS1 blockade-of-binding (BOB) ELISA to test the convalescent sera of non-flavivirus, primary DENV, secondary DENV, and ZIKV infections. Our findings indicate that primary testing using a ZIKV NS1 IgG ELISA, the test of choice for large-scale ZIKV serosurvey studies, provided relatively high sensitivity. Moreover, the confirmation of positive ELISA results using the ZIKV NS1 BOB ELISA increased average specificity to 94.59% across serum samples. The combined use of two simple ELISAs for ZIKV serosurveys and the monitoring of ZIKV infection during pregnancy can elucidate the epidemiology, pathogenesis, and complications of ZIKV in DENV-endemic areas.

7.
Trop Med Infect Dis ; 7(7)2022 Jul 12.
Article in English | MEDLINE | ID: mdl-35878144

ABSTRACT

Serology remains a useful indirect method of diagnosing tropical diseases, especially in dengue infection. However, the current literature regarding cross-reactivity between SARS-CoV-2 and dengue serology is limited and revealed conflicting results. As a means to uncover relevant serological insight involving antibody classes against SARS-CoV-2 and cross-reactivity, anti-SARS-CoV-2 IgA, IgM, and IgG ELISA, based on spike and nucleocapsid proteins, were selected for a fever-presenting tropical disease patient investigation. The study was conducted at the Faculty of Tropical Medicine during March to December 2021. The study data source comprised (i) 170 non-COVID-19 sera from 140 adults and children presenting with acute undifferentiated febrile illness and 30 healthy volunteers, and (ii) 31 COVID-19 sera from 17 RT-PCR-confirmed COVID-19 patients. Among 170 non-COVID-19 samples, 27 were false positives (15.9%), of which IgA, IgM, and IgG cross-reactive antibody classes were detected in 18 (10.6%), 9 (5.3%), and 3 (1.8%) cases, respectively. Interestingly, one case exhibited both IgA and IgM false positivity, while two cases exhibited both IgA and IgG false positivity. The false positivity rate in anti-SARS-CoV-2 IgA and IgM was reported in adults with dengue infection (11.3% and 5%) and adults with other tropical diseases (16.7% and 13.3%). The urea dissociation method applied to mitigate false positivity resulted in significantly decreased ELISA-based false and true positives. In conclusion, the analysis of antibody against SARS-CoV-2 in sera of patients with different tropical diseases showed that high IgA and IgM false positivity thus potentially limits serological assay utility in fever-presenting patients in tropical areas.

8.
PLoS Negl Trop Dis ; 16(5): e0010398, 2022 05.
Article in English | MEDLINE | ID: mdl-35587502

ABSTRACT

Most cases of dengue virus infection are mild, but severe cases can be fatal. Therefore, identification of factors associated with dengue severity is essential to improve patient outcomes and reduce mortality. The objective of this study was to assess associations between nutritional status and dengue severity among Thai children and adolescents. This retrospective cross-sectional study was based on the medical records of 355 patients with dengue treated at the Hospital for Tropical Disease (Bangkok, Thailand) from 2017 to 2019. Subjects were Thai children aged less than 18 years with dengue virus infection confirmed by positive NS1 antigen or IgM. The 1997 and 2009 World Health Organization (WHO) dengue classifications were used to define disease severity and body mass index for age while the WHO growth chart was used to classify nutritional status. The proportions of patients with dengue fever who were underweight, normal weight, and overweight were 8.8%, 61.5%, and 29.7%, respectively. The proportions of patients with dengue haemorrhagic fever (DHF) who were underweight, normal weight, and overweight were 10.2%, 66.1%, and 23.7%, respectively. The proportions of patients with non-severe dengue who were underweight, normal weight, and overweight were 8.6%, 60.9%, and 30.5%, respectively; the same proportions of patients with severe dengue were 10.5%, 67.1%, and 22.4%, respectively. Higher proportions of patients with severe plasma leakage (DHF grade III and IV) were overweight compared with those with mild plasma leakage (DHF grade I and II) (45.5% vs. 18.8%). No difference in nutritional status was observed in patients with different dengue severity.


Subject(s)
Dengue , Virus Diseases , Adolescent , Child , Cross-Sectional Studies , Dengue/complications , Dengue/epidemiology , Humans , Nutritional Status , Overweight , Retrospective Studies , Thailand/epidemiology , Thinness
9.
Viruses ; 14(3)2022 02 25.
Article in English | MEDLINE | ID: mdl-35336883

ABSTRACT

In 2016, Zika virus (ZIKV) infection was declared a public health emergency of international concern because of the neurological consequences in babies born to infected people. Because of the mild and nonspecific symptoms, serological tests are essential in epidemiological studies. However, cross-reactive antibodies between other Flaviviridae members may complicate the interpretation of results of these tests. This study investigated the seroprevalence of ZIKV infection in Samut Songkhram in central Thailand which was affected by the Zika outbreak of 2016. Three hundred and fifty volunteers aged 5-50 years in Amphawa District, Samut Songkhram, were enrolled between April 2017 and April 2018. ZIKV nonstructural protein 1 (NS1) immunoglobulin G enzyme-linked immunosorbent assay (ELISA) was used to screen serum samples collected on the first day of enrollment and after 6 and 12 months. The seroprevalence and seroconversion of ZIKV were assessed. Cases of ZIKV seroconversion were verified as evidence of ZIKV infection by NS1 blockade-of-binding ELISA and plaque reduction neutralization test (PRNT50). ZIKV seroprevalence in Amphawa was 15.1-17.8% with no significant change over the year. The total seroconversion rate throughout the year was 7/100 person-years. The ratio of asymptomatic to symptomatic infections was 4.5:1. The cases in our study confirmed the occurrence of occult ZIKV infections in the community. These undetected infections might promote the spread of ZIKV in vulnerable groups of the community.


Subject(s)
Dengue , Zika Virus Infection , Zika Virus , Antibodies, Viral , Enzyme-Linked Immunosorbent Assay/methods , Humans , Pandemics , Seroepidemiologic Studies , Serologic Tests , Thailand/epidemiology
10.
PLoS Negl Trop Dis ; 16(1): e0010021, 2022 01.
Article in English | MEDLINE | ID: mdl-34982768

ABSTRACT

BACKGROUND: To determine the seroprevalence and transmission dynamics of dengue virus (DENV), age-stratified longitudinal serological surveys were conducted in Bangphae district, Ratchaburi province, Thailand, for 3 years between April 2012 and April 2015. METHODOLOGY: The surveys enrolled 2012 healthy children and adults between 1 and 55 years-of-age, and a longitudinal serosurvey of six repeated bleeds of the same cohort of individuals was conducted every 8 months for the first 2 years (M0, M8, M16) and every half a year (M24, M30, M36) for the rest of the study period. All samples were tested using in-house indirect sandwich dengue IgG ELISA to determine DENV antibody titer, and 640 paired samples which showed rising of DENV IgG titers in paired serum were further tested using in-house neutralization assay, Plaque Reduction Neutralization Test (PRNT50). PRINCIPAL FINDINGS: When compared against the gold standard based on the results of PRNT50, sensitivity and specificity of indirect ELISA were found to be both about 85%. The overall DENV IgG positivity determined by ELISA was 74.3% in 2012 and increased to 79.4% by the final sample collection in 2015. In our study sample, more than 98% of subjects older than 25 years were found to be seropositive. Among 518 IgG negative subjects at enrollment, the seroconversion rates were measured in paired bleeds; the rates (between successive visits, approximately 6 months) ranged between 4.8% (between M16 and M24) and 14.7% (between M0 and M8). The dominant serotype of primary DENV infection cases based on seroconversion was identified from the PRNT results and it was DENV-2. CONCLUSIONS: Our study documented high levels of seroprevalence and rate of transmission. Given the importance of the serostatus and disease burden in consideration for dengue vaccine introduction, our data could be used in decision-making on implementation of various dengue control and preventive measures.


Subject(s)
Dengue/epidemiology , Seroepidemiologic Studies , Adolescent , Adult , Antibodies, Viral/blood , Child , Child, Preschool , Cohort Studies , Humans , Immunoglobulin G/blood , Incidence , Infant , Longitudinal Studies , Middle Aged , Thailand/epidemiology , Viral Plaque Assay , Young Adult
11.
J Public Health (Oxf) ; 44(1): 77-83, 2022 03 07.
Article in English | MEDLINE | ID: mdl-33993313

ABSTRACT

BACKGROUND: Undernutrition has been shown to be associated with various infectious diseases. However, the recent improvement in nutritional status and management for infectious diseases worldwide necessitates the re-evaluation of this association. METHODS: A retrospective study was conducted in children aged <14 years old with dengue, malaria or acute diarrhea who visited or were admitted to Tha Song Yang hospital, near the Thai-Myanmar border. RESULTS: Most of the patients had mild disease and most of the undernourishment was mild. The prevalence of underweight in dengue, malaria and acute diarrhea was 24.0%, 34.7% and 38.7%, respectively, and the prevalence of low height for age was 12.0%, 36.0% and 36.0%, respectively. Malaria and acute diarrhea were associated with underweight but not low height for age. Dengue was neither associated with underweight nor low height for age. CONCLUSION: Although there has been an improvement in nutritional status and health care facilities, underweight has been still prevalent in rural areas and associated with malaria and acute diarrhea. IMPLICATION: The surveillance for nutritional status should be continuously performed particularly in children with some diseases, e.g. malaria and acute diarrhea, and additional food supplementation should be provided.


Subject(s)
Dengue , Malaria , Malnutrition , Adolescent , Child , Dengue/complications , Dengue/epidemiology , Diarrhea/epidemiology , Humans , Infant , Malaria/complications , Malaria/epidemiology , Malnutrition/complications , Malnutrition/epidemiology , Myanmar/epidemiology , Nutritional Status , Prevalence , Retrospective Studies , Thailand/epidemiology , Thinness/complications
12.
Viruses ; 13(9)2021 09 10.
Article in English | MEDLINE | ID: mdl-34578383

ABSTRACT

Zika virus (ZIKV) is the mosquito-transmitted virus that the WHO declared a Public Health Emergency of International Concern in 2016 due to the consequence of microcephaly from infected pregnancies. The incidence of Zika infection has been unclear in many countries because most infected people have nonspecific febrile illnesses. This study's aim is to investigate the incidence of symptomatic Zika virus infections from the archived samples of a dengue cohort study of children in central Thailand from 2006 to 2009. We performed Zika NS1 immunoglobulin (Ig)G enzyme-linked immunosorbent assay (ELISA) screening to identify symptomatic Zika infections in paired acute/convalescent serum samples. Symptomatic Zika infections were confirmed by reverse transcription polymerase chain reactions (RT-PCR) of acute serum samples. The comparison of the Zika NS1 IgG ELISA results between acute and convalescent samples showed 290/955 (30.4%) seropositive cases. Zika RT-PCR results were positive in 28 febrile cases (15 females, 13 males). Zika RT-PCR showed that symptomatic Zika infection occurred in children aged 4-11 years in Ratchaburi province, Thailand (2007-2009, first case in April 2007), and the symptomatic Zika:dengue infection ratio was 28 Zika:394 dengue (1:14). Phylogenetic analysis showed that all Zika viruses were of Asian lineage. Zika NS1 IgG ELISA identified Zika-infected patients and showed a low Zika:dengue ratio.


Subject(s)
Dengue/epidemiology , Epidemiological Monitoring , Phylogeny , Zika Virus Infection/epidemiology , Zika Virus/genetics , Antibodies, Viral/blood , Child , Child, Preschool , Cohort Studies , Dengue Virus/pathogenicity , Female , Humans , Incidence , Male , Thailand/epidemiology , Zika Virus/classification
13.
Pathogens ; 10(5)2021 May 10.
Article in English | MEDLINE | ID: mdl-34068676

ABSTRACT

Hand, foot, and mouth disease (HFMD) is a contagious childhood illness and annually affects millions of children aged less than 5 years across the Asia-Pacific region. HFMD transmission mainly occurs through direct contact (person-to-person) and indirect contact with contaminated surfaces and objects. Therefore, public health measures to reduce the spread of HFMD in kindergartens and daycare centers are essential. Based on the guidelines by the Department of Disease Control, a school closure policy for HFMD outbreaks wherein every school in Thailand must close when several HFMD classrooms (more than two cases in each classroom) are encountered within a week, was implemented, although without strong supporting evidence. We therefore conducted a prospective cohort study of children attending five kindergartens during 2019 and 2020. We used molecular genetic techniques to investigate the characteristics of the spreading patterns of HFMD in a school-based setting in Bangkok, Thailand. These analyses identified 22 index cases of HFMD (symptomatic infections) and 25 cases of enterovirus-positive asymptomatic contacts (24 students and one teacher). Enterovirus (EV) A71 was the most common enterovirus detected, and most of the infected persons (8/12) developed symptoms. Other enteroviruses included coxsackieviruses (CVs) A4, CV-A6, CV-A9, and CV-A10 as well as echovirus. The pattern of the spread of HFMD showed that 45% of the subsequent enteroviruses detected in each outbreak possessed the same serotype as the first index case. Moreover, we found a phylogenetic relationship among enteroviruses detected among contact and index cases in the same kindergarten. These findings confirm the benefit of molecular genetic assays to acquire accurate data to support school closure policies designed to control HFMD infections.

14.
Int J Infect Dis ; 107: 271-277, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33991681

ABSTRACT

OBJECTIVES: This study aimed to evaluate a microfluidic paper-based analytical device (DEN-NS1-PAD) based on a rapid NS1 antigen test for diagnosing dengue at the point of care. METHODS: 219 serum samples from suspected dengue cases were tested with the developed DEN-NS1-PAD and commercial RDT by SD BIOLINE. The results were compared with the nested-PCR results. RESULTS: The limit of detection of DEN-NS1-PAD was 0.78 ng mL-1. It showed 88.89% sensitivity, 86.67% specificity, and a substantial agreement correlation (κ = 0.7522) compared with nested-PCR. In contrast, SD BIOLINE for NS1 (SD-NS1) detection showed 87.88% sensitivity, 90.00% specificity, and had a substantial agreement correlation with nested-PCR (κ = 0.7788). CONCLUSIONS: DEN-NS1-PAD is a valuable tool for diagnosing DENV infections, especially for diagnosed patients with early acute phase samples with high viral load. DEN-NS1-PAD has better sensitivity than SD-NS1 but less specificity.


Subject(s)
Antigens, Viral/analysis , Dengue Virus/immunology , Fever/diagnosis , Fever/virology , Paper , Point-of-Care Testing , Antigens, Viral/immunology , Humans , Male , Middle Aged , Time Factors
15.
Anal Bioanal Chem ; 412(12): 2915-2925, 2020 May.
Article in English | MEDLINE | ID: mdl-32166444

ABSTRACT

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 20-30 min. 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 74.8 ng mL-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.


Subject(s)
Antibodies, Viral/blood , Antigens, Viral/immunology , Dengue Virus/isolation & purification , Dengue/diagnosis , Microfluidics/instrumentation , Microfluidics/methods , Viral Nonstructural Proteins/blood , Case-Control Studies , Child , Dengue/metabolism , Dengue/virology , Humans , Immunoassay , Viral Nonstructural Proteins/immunology
16.
Jpn J Infect Dis ; 73(2): 140-147, 2020 Mar 24.
Article in English | MEDLINE | ID: mdl-31787738

ABSTRACT

Dengue is one of the most serious mosquito-borne viral diseases occurring in humans. To combat the complexity of 4 antigenically distinct serotypes, the ideal vaccine for dengue should be able to stimulate cross-neutralizing antibodies. Recently, genetics-based immune responses have been studied to guide vaccine design against several viral pathogens. Despite a recent approval of dengue vaccine, information on genetics-based immune responses against dengue virus (DENV) is still limited. Consequently, we aimed to determine the profiles of immunoglobulin heavy chain genes from DENV2 infected patients. The immunoglobulin heavy chain variable region genes (IGHV) were amplified from peripheral blood mononuclear cells of DENV2 secondary infected patients in the acute, convalescence, and recovery phases. Antibody heavy chain genes were sequenced using next-generation sequencing, and analyzed to identify correlations with neutralizing and enhancing activities of the serum samples. IGHV1-69, 3-23, and 3-30 were frequently discovered in our Thai DENV2 infected patients. Our findings provide new data on the human B cell response during secondary DENV2 infections in Thai dengue patients that offer supportive information for dengue vaccine design and therapeutics development.


Subject(s)
Coinfection/immunology , Dengue/immunology , Genes, Immunoglobulin Heavy Chain/genetics , Adolescent , Adult , Antibodies, Viral/blood , B-Lymphocytes/immunology , Coinfection/genetics , Coinfection/virology , Dengue/genetics , Dengue/virology , Dengue Virus , High-Throughput Nucleotide Sequencing , Humans , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/virology , Sequence Analysis, DNA , Serogroup , Young Adult
17.
Int Health ; 10(5): 349-355, 2018 09 01.
Article in English | MEDLINE | ID: mdl-29912451

ABSTRACT

Background: This study focused on evaluating the fumigation scheme and identifying problems encountered during the operation in the Bangkok Metropolitan Administration area. Methods: Ten district health officers working in different fumigation teams of the dengue outbreak control programme around Bangkok had participated in an in-depth interview. Five predetermined themes, including (i) dengue surveillance and control strategy, (ii) quality and availability of equipment, (iii) delays, (iv) human resources, and (v) area coverage, and other emerging themes were addressed during the interviews. Results: Although the staff seemed to know the operation protocol of the dengue surveillance and control programmes well, they encountered some difficulties in accessing households for proper spraying, and a lack of human and material resources, especially during an outbreak. Other emerging themes concerned inefficient communications among the sectors from hospital to district offices, leading to inaccurate or missing patient addresses for spraying, and the lack of community networks and public cooperation for the dengue control programmes. Conclusions: The findings suggest that coordination among the relevant health sectors to acquire accurate and timely information about dengue cases is essential. Involving community networks should help to improve public engagement with and participation in the surveillance and outbreak control programmes.


Subject(s)
Community Networks/organization & administration , Dengue/prevention & control , Fumigation/statistics & numerical data , Insecticides/administration & dosage , Mosquito Control/methods , Government Programs , Humans , Thailand , Workforce
18.
Nephron ; 139(1): 70-82, 2018.
Article in English | MEDLINE | ID: mdl-29402790

ABSTRACT

BACKGROUND: Previous studies showed that ceftriaxone can cause acute kidney injury (AKI) in the pediatric population. This study proposed a cellular model of crystalline nephropathy in ceftriaxone-associated AKI and explored the related pathophysiology by using a proteomic approach. METHODS: Ceftriaxone was crystallized with calcium in artificial urine. Madin-Darby Canine Kidney (MDCK) cells, a model of distal renal tubular cell, were cultured in the absence (untreated control) or presence of ceftriaxone crystals for 48-h (n = 5 each). MDCK cells were harvested and subsequently analyzed by proteomic analysis. Protein bioinformatics (i.e., STRING and Reactome) was used to predict functional alterations, and subsequently validated by Western blotting and cellular studies. p < 0.05 was considered statistically significant. RESULTS: Phase-contrast microscopy showed increased intracellular vesiculation and cell enlargement as a result of ceftriaxone crystal exposure. Proteome analysis revealed a total of 20 altered proteins (14 increased, 5 decreased and 1 absent) in ceftriaxone crystal-treated MDCK cells as compared to untreated cells (p < 0.05). Protein bioinformatics and validation studies supported heat stress response mediated by heat shock protein 70 (Hsp70) and downregulation of annexin A1 as the proposed pathophysiology of crystalline nephropathy in ceftriaxone-associated AKI, in which impaired proliferation and wound healing of crystal-induced distal tubular cells were outcomes. CONCLUSIONS: This study, for the first time, used the in vitro model of crystalline nephropathy to investigate the underlying pathophysiology of ceftriaxone-associated AKI, which should be investigated in vivo for potential clinical benefits in the future.


Subject(s)
Acute Kidney Injury/chemically induced , Acute Kidney Injury/physiopathology , Anti-Infective Agents/adverse effects , Ceftriaxone/adverse effects , Acute Kidney Injury/pathology , Animals , Annexin A1/biosynthesis , Calcium/chemistry , Cell Proliferation , Crystallization , Dogs , HSP70 Heat-Shock Proteins/metabolism , Kidney Tubules, Distal/pathology , Madin Darby Canine Kidney Cells , Proteomics
19.
Proteomics Clin Appl ; 11(7-8)2017 07.
Article in English | MEDLINE | ID: mdl-28195677

ABSTRACT

Plasma is a rich source of biomarkers with clinical relevance. However, the wide dynamic range of protein concentration hinders the detection of low abundance proteins. Plasma prefractionation methods serve as indispensable tools to reduce plasma complexity, allowing the opportunity to explore tissue-derived proteins which leak into the circulation. This review summarizes common approaches in plasma prefractionation methods for proteomic analysis and then discusses some considerations in plasma prefractionation for clinical applications, reviewing some examples of its use in clinical situations.


Subject(s)
Analytic Sample Preparation Methods/methods , Blood Proteins/isolation & purification , Blood Proteins/metabolism , Chemical Fractionation/methods , Proteomics/methods , Animals , Humans
20.
Asian Pac J Trop Med ; 10(1): 11-14, 2017 Jan.
Article in English | MEDLINE | ID: mdl-28107858

ABSTRACT

Dengue virus (DENV) is a mosquito-transmitted virus that is expanding across the world. The incidence of dengue infection, especially severe disease, has been increasing. DENV consist of 4 serotypes of single stranded RNA viruses (D1-D4) in the genus Flavivirus, family Flaviviridae. Majority of dengue infections are asymptomatic cases, which cause difficulty in disease control and are important in dengue surveillance. There is still no gold standard to diagnose asymptomatic dengue infection. Plaque reduction neutralization test (PRNT) has been developed for many purposes such as immunological study, clinical study, vaccine trial and is currently the most sensitive and specific method for serological surveillance. However, PRNT shows some degree of cross reaction among different dengue serotypes especially secondary dengue infection cases and to other flaviviruses. Moreover, various modification since the beginning make PRNT lack of inter-laboratory standardization which is an important issue. This paper discusses the important of asymptomatic dengue infection and its diagnostic method.

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