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1.
PeerJ ; 12: e16773, 2024.
Article in English | MEDLINE | ID: mdl-38313031

ABSTRACT

Background: Human opisthorchiasis is a dangerous infectious chronic disease distributed in many Asian areas in the water-basins of large rivers, Siberia, and Europe. The gold standard for human opisthorchiasis laboratory diagnosis is the routine examination of Opisthorchis spp. eggs under a microscope. Manual detection is laborious, time-consuming, and dependent on the microscopist's abilities and expertise. Automatic screening of Opisthorchis spp. eggs with deep learning techniques is a useful diagnostic aid. Methods: Herein, we propose a convolutional neural network (CNN) for classifying and automatically detecting O. viverrini eggs from digitized images. The image data acquisition was acquired from infected human feces and was processed using the gold standard formalin ethyl acetate concentration technique, and then captured under the microscope digital camera at 400x. Microscopic images containing artifacts and O.viverrini egg were augmented using image rotation, filtering, noising, and sharpening techniques. This augmentation increased the image dataset from 1 time to 36 times in preparation for the training and validation step. Furthermore, the overall dataset was subdivided into a training-validation and test set at an 80:20 ratio, trained with a five-fold cross-validation to test model stability. For model training, we customized a CNN for image classification. An object detection method was proposed using a patch search algorithm to detect eggs and their locations. A performance matrix was used to evaluate model efficiency after training and IoU analysis for object detection. Results: The proposed model, initially trained on non-augmented data of artifacts (class 0) and O. viverrini eggs (class 1), showed limited performance with 50.0% accuracy, 25.0% precision, 50.0% recall, and a 33.0% F1-score. After implementing data augmentation, the model significantly improved, reaching 100% accuracy, precision, recall, and F1-score. Stability assessments using 5-fold cross-validation indicated better stability with augmented data, evidenced by an ROC-AUC metric improvement from 0.5 to 1.00. Compared to other models such as ResNet50, InceptionV3, VGG16, DenseNet121, and Xception, the proposed model, with a smaller file size of 2.7 MB, showed comparable perfect performance. In object detection, the augmented data-trained model achieved an IoU score over 0.5 in 139 out of 148 images, with an average IoU of 0.6947. Conclusion: This study demonstrated the successful application of CNN in classifying and automating the detection of O. viverrini eggs in human stool samples. Our CNN model's performance metrics and true positive detection rates were outstanding. This innovative application of deep learning can automate and improve diagnostic precision, speed, and efficiency, particularly in regions where O. viverrini infections are prevalent, thereby possibly improving infection sustainable control and treatment program.


Subject(s)
Opisthorchiasis , Opisthorchis , Animals , Humans , Opisthorchiasis/diagnosis , Feces , Research Design , Neural Networks, Computer
2.
Parasitol Res ; 123(2): 115, 2024 Jan 29.
Article in English | MEDLINE | ID: mdl-38285129

ABSTRACT

Hookworm infections remain a significant public health concern in tropical and subtropical regions, including Thailand. This study investigated the species and genetic diversity of hookworm infections in domestic dogs from northeastern Thailand. The molecular analysis focused on amplifying and sequencing specific regions of ribosomal RNA genes (ITS1-5.8S-ITS2 region) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene in hookworm larvae recovered from 21 domestic dog stool samples. Among 21 larvae (one larva per infected dog) analyzed, 14 had sequences identical to Ancylostoma caninum, and 7 showed sequences almost identical to Ancylostoma ceylanicum. Phylogenetic analysis of cox1 sequences placed A. caninum and A. ceylanicum in separate clades. The median-joining network of A. caninum cox1 sequences from Thailand showed high haplotype diversity and belonged to the same cluster as sequences from Australia while forming separate clusters from those of A. caninum samples from the USA. The available published A. ceylanicum cox1 sequences (n = 33), in combination with seven sequences in the present study, represented 15 haplotypes distributed among three clusters. Interestingly, A. ceylanicum sequences from dogs and humans shared the same haplotypes. These findings are crucial for recognizing the potential for zoonotic transmission, highlighting the necessity for targeted control measures, and increasing awareness among pet owners and healthcare professionals to mitigate the risk of hookworm transmission to humans.


Subject(s)
Ancylostomatoidea , Hookworm Infections , Humans , Animals , Dogs , Ancylostomatoidea/genetics , Phylogeny , Thailand/epidemiology , Zoonoses/epidemiology , Hookworm Infections/epidemiology , Hookworm Infections/veterinary , Ancylostoma/genetics , Larva , Genetic Variation
3.
Parasite ; 30: 60, 2023.
Article in English | MEDLINE | ID: mdl-38099622

ABSTRACT

Strongyloidiasis is a neglected tropical disease that can cause fatal complications due to hyperinfection and disseminated strongyloidiasis in immunocompromised patients. We used two Strongyloides stercoralis recombinant antigenic proteins, L3NieAg.01 (NIE) and IgG-immunoreactive antigen (SsIR), to develop the recombinant antigen-based immunochromatography test (ICT) kit. We constructed and compared kits using either the NIE (NIE ICT kit) or the SsIR (SsIR ICT kit) antigens and a kit using a mixture of both (NIE-SsIR ICT kit) for detection of anti-Strongyloides IgG antibody in human serum samples. Serum samples from normal healthy individuals (Group I, n = 40), proven strongyloidiasis patients (Group II, n = 100), and those with other parasitic infections (Group III, n = 154) were evaluated. Sensitivity and specificity were 81.0% and 84.0% for the NIE ICT kit, 89.0% and 83.5% for the SsIR ICT kit, and 95.0% and 90.2% for the NIE-SsIR ICT kit, respectively. The NIE-SsIR ICT kit provided the best diagnostic results; it can supplement stool examination for clinical diagnosis and can be used to screen for asymptomatic S. stercoralis infection in people at risk in endemic areas. The NIE-SsIR ICT kit can also be used in large-scale sero-epidemiological investigations in endemic areas without the need for additional facilities or ancillary supplies.


Title: Amélioration de la sensibilité diagnostique de l'anguillulose humaine grâce à l'immunochromatographie à base d'antigènes recombinants mixtes sur le lieu d'intervention. Abstract: La strongyloïdose est une maladie tropicale négligée qui peut entraîner des complications mortelles dues à une hyperinfection et à une strongyloïdose disséminée chez les patients immunodéprimés. Nous avons utilisé deux protéines antigéniques recombinantes de Strongyloides stercoralis, L3NieAg.01 (NIE) et l'antigène immunoréactif IgG (SsIR), pour développer un kit de test d'immunochromatographie (TIC) basé sur les antigènes recombinants. Nous avons construit et comparé des kits utilisant les antigènes NIE (kit NIE ICT), SsIR (kit SsIR ICT) ou un mélange des deux (kit NIE-SsIR ICT) pour la détection des anticorps IgG anti-Strongyloides dans des échantillons de sérum humain. Des échantillons de sérum provenant d'individus normaux en bonne santé (groupe I, n = 40), de patients atteints d'anguillulose avérée (groupe II, n = 100) et de patients atteints d'autres infections parasitaires (groupe III, n = 154) ont été évalués. La sensibilité et la spécificité étaient respectivement de 81,0 % et 84,0 % pour le kit NIE ICT, 89,0 % et 83,5 % pour le kit SsIR ICT, et 95,0 % et 90,2 % pour le kit NIE-SsIR ICT. Le kit NIE-SsIR ICT a fourni les meilleurs résultats de diagnostic ; il peut compléter l'examen des selles pour le diagnostic clinique et peut être utilisé pour dépister une infection asymptomatique à S. stercoralis chez les personnes à risque dans les zones d'endémie. Le kit NIE-SsIR ICT peut également être utilisé dans des enquêtes séro-épidémiologiques à grande échelle dans les zones endémiques sans nécessiter d'installations supplémentaires ou de fournitures auxiliaires.


Subject(s)
Strongyloides stercoralis , Strongyloidiasis , Animals , Humans , Strongyloidiasis/diagnosis , Strongyloidiasis/parasitology , Point-of-Care Systems , Antibodies, Helminth , Sensitivity and Specificity , Chromatography, Affinity/methods
4.
Animals (Basel) ; 13(20)2023 Oct 13.
Article in English | MEDLINE | ID: mdl-37893924

ABSTRACT

Opisthorchiasis and clonorchiasis are prevalent in Southeast and Far-East Asia, which are caused by the group 1 carcinogenic liver flukes Opisthorchis viverrini sensu lato and Clonorchis sinensis infection. There have been comprehensive investigations of systematics and genetic variation of these liver flukes. Previous studies have shown that O. viverrini is a species complex, called "O. viverrini sensu lato". More comprehensive investigations of molecular systematics and population genetics of each of the species that make up the species complex are required. Thus, other polymorphic genetic markers need to be developed. Therefore, this study aimed to characterize the intron regions of taurocyamine kinase gene (TK) to examine the genetic variation and population genetics of O. viverrini and C. sinensis collected from different geographical isolates and from a range of animal hosts. We screened seven intron regions embedded in TK. Of these, we selected an intron 5 of domain 1 (TkD1Int5) region to investigate the genetic variation and population genetics of theses liver flukes. The high nucleotide and haplotype diversity of TkD1Int5 was detected in O. viverrine. Heterozygosity with several insertion/deletion (indel) regions were detected in TkD1Int5 of the O. viverrine samples, whereas only an indel nucleotide was detected in one C. sinensis sample. Several O. viverrine samples contained three different haplotypes within a particular heterozygous sample. There were no genetic differences between C. sinensis isolated from various animal host. Heterozygous patterns specifically detected in humans was observed in C. sinensis. Thus, TkD1Int5 is a high polymorphic genetic marker, which could be an alternative marker for further population genetic investigations of these carcinogenic liver flukes and other related species from a wide geographical distribution and variety of animal hosts.

5.
Food Waterborne Parasitol ; 33: e00211, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37868190

ABSTRACT

Human cysticercosis is a life-threatening zoonotic disease caused by infection with larvae (cysticerci) of the pork tapeworm, Taenia solium. This can affect the nervous system causing chronic headache and intracranial hypertension, potentially leading to epileptic seizures and paralysis. The disease is found in developing countries, especially in Southeast and South Asia, Sub-Saharan Africa, and Central and South America where porcine cysticercosis is endemic and people have a habit of eating undercooked pork. An immunochromatography-based test (ICT) kit, using T. solium cyst fluid as antigen, was manufactured to detect anti-T. solium IgG antibodies in human serum. To evaluate the kit, we used 187 serum samples including 24 from proven/confirmed cysticercosis cases, 133 from cases with other parasitosis and 30 healthy controls. Diagnostic efficiencies were calculated. The sensitivity, specificity, and accuracy were 83.3%, 92.0%, and 90.9%, respectively. Moreover, the ICT was positive before treatment but became negative after treatment, implying that this kit is also useful for follow-up monitoring post-treatment. In conclusion, we have successfully developed and present preliminary evaluation of an easy-to-handle rapid diagnostic tool for human cysticercosis in the form of an ICT platform using as antigen fluid from T. solium cysticerci.

6.
Infect Genet Evol ; 115: 105511, 2023 11.
Article in English | MEDLINE | ID: mdl-37820843

ABSTRACT

Ticks are vectors of a variety of pathogens that can infect humans and animals. Ticks also harbor non-pathogenic microbiota. This study characterized the microbiota of the ticks infesting beef cattle in Thailand. Two species of ticks; Rhipicephalus microplus (n = 15) and Haemaphysalis bispinosa (n = 5), were collected in seven provinces in northeastern Thailand. Microbial community profile of ticks was examined based on sequences of the V3-V4 region of 16S rRNA gene. Proteobacteria (Pseudomonadota) was the most abundant phylum, followed by Firmicutes (Bacillota), and Actinobacteriota. Coxiella-like endosymbiont was the most abundant bacterial taxon overall (49% of sequence reads), followed by Anaplasma (8.5%), Corynebacterium (5.5%), Ehrlichia (3.9%), and Castellaniella (3.4%). Co-infections of the pathogenic bacteria Ehrlichia and Anaplasma were detected in 19/20 (95%) female ticks. The tick with the lowest number of bacteria had the lowest abundance of the Coxiella-like endosymbiont, and the pathogenic bacteria Anaplasma and Ehrlichia were absent. This study provides baseline information of the microbiota of cattle ticks in northeastern Thailand, suggesting that ticks carry a few dominant bacterial taxa that are primarily non-pathogenic but can co-occur with pathogenic microorganisms. The information obtained is useful for monitoring disease outbreaks in the future and informing prevention and control strategies against cattle tick-borne diseases.


Subject(s)
Microbiota , Rhipicephalus , Rickettsia , Tick-Borne Diseases , Animals , Humans , Female , Male , RNA, Ribosomal, 16S/genetics , Thailand/epidemiology , Bacteria/genetics , Rhipicephalus/genetics , Ehrlichia/genetics , Tick-Borne Diseases/epidemiology , Anaplasma/genetics , Microbiota/genetics , Rickettsia/genetics
7.
PLoS One ; 18(8): e0289733, 2023.
Article in English | MEDLINE | ID: mdl-37590198

ABSTRACT

Bacterial content of mosquitoes has given rise to the development of innovative tools that influence and seek to control malaria transmission. This study identified the bacterial microbiota in field-collected female adults of the Anopheles hyrcanus group and three Anopheles species, Anopheles nivipes, Anopheles philippinensis, and Anopheles vagus, from an endemic area in the southeastern part of Ubon Ratchathani Province, northeastern Thailand, near the Lao PDR-Cambodia-Thailand border. A total of 17 DNA libraries were generated from pooled female Anopheles abdomen samples (10 abdomens/ sample). The mosquito microbiota was characterized through the analysis of DNA sequences from the V3-V4 regions of the 16S rRNA gene, and data were analyzed in QIIME2. A total of 3,442 bacterial ASVs were obtained, revealing differences in the microbiota both within the same species/group and between different species/group. Statistical difference in alpha diversity was observed between An. hyrcanus group and An. vagus and between An. nivipes and An. vagus, and beta diversity analyses showed that the bacterial community of An. vagus was the most dissimilar from other species. The most abundant bacteria belonged to the Proteobacteria phylum (48%-75%) in which Pseudomonas, Serratia, and Pantoea were predominant genera among four Anopheles species/group. However, the most significantly abundant genus observed in each Anopheles species/group was as follows: Staphylococcus in the An. hyrcanus group, Pantoea in the An. nivipes, Rosenbergiella in An. philippinensis, and Pseudomonas in An. vagus. Particularly, Pseudomonas sp. was highly abundant in all Anopheles species except An. nivipes. The present study provides the first study on the microbiota of four potential malaria vectors as a starting step towards understanding the role of the microbiota on mosquito biology and ultimately the development of potential tools for malaria control.


Subject(s)
Anopheles , Malaria , Pantoea , Animals , Female , RNA, Ribosomal, 16S/genetics , Thailand/epidemiology , Mosquito Vectors , Malaria/epidemiology , Pseudomonas
8.
Front Microbiol ; 14: 1052352, 2023.
Article in English | MEDLINE | ID: mdl-37032902

ABSTRACT

Infectious diseases caused by filarial nematodes are major health problems for humans and animals globally. Current treatment using anti-helminthic drugs requires a long treatment period and is only effective against the microfilarial stage. Most species of filarial nematodes harbor a specific strain of Wolbachia bacteria, which are essential for the survival, development, and reproduction of the nematodes. This parasite-bacteria obligate symbiosis offers a new angle for the cure of filariasis. In this study, we utilized publicly available genome data and putative protein sequences from seven filarial nematode species and their symbiotic Wolbachia to screen for protein-protein interactions that could be a novel target against multiple filarial nematode species. Genome-wide in silico screening was performed to predict molecular interactions based on co-evolutionary signals. We identified over 8,000 pairs of gene families that show evidence of co-evolution based on high correlation score and low false discovery rate (FDR) between gene families and obtained a candidate list that may be keys in filarial nematode-Wolbachia interactions. Functional analysis was conducted on these top-scoring pairs, revealing biological processes related to various signaling processes, adult lifespan, developmental control, lipid and nucleotide metabolism, and RNA modification. Furthermore, network analysis of the top-scoring genes with multiple co-evolving pairs suggests candidate genes in both Wolbachia and the nematode that may play crucial roles at the center of multi-gene networks. A number of the top-scoring genes matched well to known drug targets, suggesting a promising drug-repurposing strategy that could be applicable against multiple filarial nematode species.

9.
Trop Dis Travel Med Vaccines ; 9(1): 2, 2023 Feb 10.
Article in English | MEDLINE | ID: mdl-36759878

ABSTRACT

BACKGROUND: Rat lung worm disease (RLWD) has several clinical forms including eosinophilic meningitis (EOM) and two severe forms, eosinophilic meningoencephalitis (EOME) and eosinophilic radiculomyelitis (EORM). It remains unclear whether transmission sources are associated with severe forms of RLWD. This study aimed to evaluate if transmission factors are related to the severity of RLWD among travelers by using a scoping review of case reports. METHODS: This was a review using five databases to retrieve case reports and case series of travelers with RLWD. Clinical data and transmission sources of reported cases diagnosed as RLWD were retrieved. The outcome of the study was occurrence of severe forms of RLWD defined as EOME, EORM, and combined EOME/EORM. RESULTS: We retrieved 1,326 articles from five databases and 31 articles were included in the analysis. There were 84 cases eligible from 15 countries. Four cases were excluded. Seventy cases were in EOM group and 10 cases had EOME or EORM. Compared with the EOM group, the EOME, EORM, and combination EOME/EORM group had similar age, sex, and risk factors of consumptions of apple snails, shrimp and prawn, and salad/vegetables. The EOME group had higher proportion of consumption of African snails than the EOM group (60% vs 13.8%). However, only one study reported the consumption of African snails and the heterogeneity between studies and the small sample size impeded direct comparisons between groups. CONCLUSIONS: RLWD in travelers can be found in most continents and mostly get infected from endemic countries of RLWD. Further studies are required to evaluate the association between transmission vectors and severity of RLWD.

10.
Insects ; 14(2)2023 Feb 13.
Article in English | MEDLINE | ID: mdl-36835753

ABSTRACT

Bacterial content in mosquito larvae and adults is altered by dynamic interactions during life and varies substantially in variety and composition depending on mosquito biology and ecology. This study aimed to identify the microbiota in Aedes aegypti and Aedes albopictus and in water from their breeding sites in northeastern Thailand, a dengue-endemic area. Bacterial diversity in field-collected aquatic larvae and subsequently emerged adults of both species from several locations were examined. The microbiota was characterized based on analysis of DNA sequences from the V3-V4 region of the 16S rRNA gene and exhibited changes during development, from the mosquito larval stage to the adult stage. Aedes aegypti contained a significantly higher number of bacterial genera than did Ae. albopictus, except for the genus Wolbachia, which was present at significantly higher frequencies in male Ae. albopictus (p < 0.05). Our findings also indicate likely transstadial transmission from larva to adult and give better understanding of the microbial diversity in these mosquitoes, informing future control programs against mosquito-borne diseases.

11.
Infect Genet Evol ; 107: 105394, 2023 01.
Article in English | MEDLINE | ID: mdl-36502740

ABSTRACT

Genetic diversity, genetic structure and demographic history of the ticks infesting beef cattle in Thailand were examined based on mitochondrial cytochrome c oxidase I (cox1) sequences. Tick samples were collected in 12 provinces in upper-northeastern Thailand. Three species were found; Rhipicephalus microplus, R. sanguineus, and Haemaphysalis bispinosa. Of these, R. microplus was by far the most abundant species in beef cattle and was widely distributed throughout the area. No cox1 sequence variation was found in the R. sanguineus or H. bispinosa specimens collected. Low nucleotide diversity but high haplotype diversity was observed in R. microplus. All collected R. microplus specimens belonged to lineage A. Mismatch-distribution analysis, as well as Tajima's D and Fu's Fs tests, provided evidence of recent demographic expansion. A subsample of tick specimens was investigated for presence of Anaplasma and Ehrlichia using a fragment of the 16S rRNA gene. Three species of Anaplasma were detected from R. microplus; Anaplasma marginale (19.08%), Anaplasma platys (1.97%) and unidentified Anaplasma strain (0.66%). The infection rate of Ehrlichia was 7.24% (two ticks were infected with E. minasensis (1.97%) and eight with an unidentified Ehrlichia strain (5.26%). No infections were found in R. sanguineus or H. bispinosa. This is the first report of A. platys and E. minasensis in cattle ticks in Thailand, providing information for future epidemiological surveys and control strategies in this region.


Subject(s)
Ixodidae , Rhipicephalus , Tick Infestations , Animals , Cattle , Ehrlichia/genetics , Ixodidae/genetics , Thailand/epidemiology , RNA, Ribosomal, 16S/genetics , Anaplasma/genetics , Rhipicephalus/genetics , Genetic Variation , Tick Infestations/epidemiology , Tick Infestations/veterinary
12.
PLoS One ; 17(12): e0279766, 2022.
Article in English | MEDLINE | ID: mdl-36584127

ABSTRACT

Human gastrointestinal helminthic infections have a direct and/or indirect effect on the composition of the host gut microbial flora. Here, we investigated the effect of infection with a soil-transmitted intestinal nematode, Strongyloides stercoralis, on the gut microbiota of the human host. We also investigated whether composition of the microbiota in infected persons might vary across endemic regions. Fecal samples were obtained from volunteers from two areas endemic for strongyloidiasis, Khon Kaen Province in northeastern Thailand and Nakhon Si Thammarat Province in southern Thailand. Samples from Khon Kaen were from infected (SsNE) and uninfected (NegNE) individuals. Similarly, samples from the latter province were from infected (SsST) and uninfected (NegST) individuals. DNA sequences of the V3-V4 regions of the bacterial 16S rRNA gene were obtained from the fecal samples. No statistical difference in alpha diversity between groups in terms of richness or diversity were found. Statistical difference in beta diversity was observed only between NegNE and NegST. Some significant differences in species abundance were noted between geographical isolates. The SsNE group had a higher abundance of Tetragenococcus holophilus than did the SsST group, whereas Bradyrhizobium sp. was less abundant in the SsNE than the SsST group. For the uninfected groups, the NegNE had a higher abundance of T. holophilus than the NegST group. Our data showed that S. stercoralis infection leads to only minor alterations in the relative abundance of individual bacterial species in the human gut: no detectable effect was observed on community structure and diversity.


Subject(s)
Gastrointestinal Microbiome , Strongyloides stercoralis , Strongyloidiasis , Animals , Humans , Strongyloidiasis/epidemiology , Gastrointestinal Microbiome/genetics , Thailand/epidemiology , RNA, Ribosomal, 16S/genetics , Strongyloides stercoralis/genetics , Feces/microbiology
13.
PLoS One ; 17(12): e0279754, 2022.
Article in English | MEDLINE | ID: mdl-36584196

ABSTRACT

Intestinal parasitic infections (IPIs) remain a public-health problem worldwide, including in countries of the Lower Mekong subregion. Increases in human migration from neighboring countries might cause reemerging parasitic infections, leading to spread of parasites in the landscape. Here, we conducted a cross-sectional study to identify the prevalence of IPIs in migrant workers from Myanmar, Lao PDR, and Cambodia who were dwelling in Nakhon Ratchasima Province, northeastern Thailand. The identification of Strongyloides species and genetic differentiation of worms from migrant workers with different countries of origin was also assessed. Fresh stool samples were collected from 338 migrant workers and examined for evidence of IPIs using agar plate culture (APC) and the formalin-ethyl acetate concentration technique (FECT). Among those nine samples positive for nematodes by APC, the Strongyloides or hookworm species present was confirmed using the polymerase chain reaction (PCR) followed by DNA sequencing. This revealed eight cases of Strongyloides stercoralis infection and one of Necator americanus. Fifty-one out of 338 individuals (15.09%) were positive for IPIs using FECT and APC. Eggs of Opisthorchis-like flukes were the most common parasite (11.83% of samples), followed by S. stercoralis (2.37%), Entamoeba coli (1.50%), hookworm (0.89%), Taenia sp. (0.60%) and Hymenolepis nana (0.30%). The genetic differentiation of S. stercoralis recovered from migrant workers with different countries of origin was analyzed. Specimens of S. stercoralis isolated from workers from Lao PDR, Cambodia and Myanmar were genetically similar to those sequenced from Thailand. However, there were population-genetic differences between S. stercoralis from these Southeast Asian countries and other regions of the world. This study demonstrated that IPIs were prevalent in migrant workers in the northeastern region of Thailand. Our findings provided molecular confirmation of the presence of S. stercoralis and explored the genetic differentiation of S. stercoralis from those infected migrant workers. An effective anti-parasitic drug should be provided for migrant workers and its administration enforced.


Subject(s)
Hookworm Infections , Intestinal Diseases, Parasitic , Parasites , Strongyloides stercoralis , Strongyloidiasis , Transients and Migrants , Humans , Animals , Thailand/epidemiology , Cambodia/epidemiology , Prevalence , Laos/epidemiology , Myanmar/epidemiology , Cross-Sectional Studies , Intestinal Diseases, Parasitic/epidemiology , Intestinal Diseases, Parasitic/parasitology , Strongyloidiasis/epidemiology , Strongyloidiasis/parasitology , Hookworm Infections/epidemiology , Ancylostomatoidea , Feces/parasitology
14.
PLoS One ; 17(10): e0275362, 2022.
Article in English | MEDLINE | ID: mdl-36227878

ABSTRACT

Cholangiocarcinoma (CCA) is highly prevalent in the northeastern region of Thailand. Current diagnostic methods for CCA are often expensive, time-consuming, and require medical professionals. Thus, there is a need for a simple and low-cost CCA screening method. This work developed a rapid label-free technique by Raman spectroscopy combined with the multivariate statistical methods of principal component analysis and linear discriminant analysis (PCA-LDA), aiming to analyze and classify between CCA (n = 30) and healthy (n = 30) serum specimens. The model's classification performance was validated using k-fold cross validation (k = 5). Serum levels of cholesterol (548, 700 cm-1), tryptophan (878 cm-1), and amide III (1248,1265 cm-1) were found to be statistically significantly higher in the CCA patients, whereas serum beta-carotene (1158, 1524 cm-1) levels were significantly lower. The peak heights of these identified Raman marker bands were input into an LDA model, achieving a cross-validated diagnostic sensitivity and specificity of 71.33% and 90.00% in distinguishing the CCA from healthy specimens. The PCA-LDA technique provided a higher cross-validated sensitivity and specificity of 86.67% and 96.67%. To conclude, this work demonstrated the feasibility of using Raman spectroscopy combined with PCA-LDA as a helpful tool for cholangiocarcinoma serum-based screening.


Subject(s)
Bile Duct Neoplasms , Cholangiocarcinoma , Amides , Bile Duct Neoplasms/diagnosis , Bile Ducts, Intrahepatic , Cholangiocarcinoma/diagnosis , Discriminant Analysis , Humans , Principal Component Analysis , Spectrum Analysis, Raman/methods , Tryptophan , beta Carotene
15.
PeerJ ; 10: e14085, 2022.
Article in English | MEDLINE | ID: mdl-36196402

ABSTRACT

Background: Amebic liver abscess (ALA) caused by Entamoeba histolytica is usually diagnosed based on its clinical symptoms, medical imaging abnormalities of the liver, and serological tests, the most common being the enzyme-linked immunosorbent assay (ELISA). For more than three decades, no investigation has evaluated the diagnostic performance of immunoglobulin G (IgG) subclasses in the serodiagnosis of ALA. Herein, we assessed the efficiencies of anti-amebic IgG and IgG subclasses for diagnosing ALA. Methods: A serological ELISA-based test was performed to assess its diagnostic performance using a total of 330 serum samples from ALA patients (n = 14), healthy individuals (n = 40), and patients with other diseases (n = 276). Results: ELISA targeting the total IgG antibody to E. histolytica antigen exhibited 100% sensitivity 95% CI [76.8-100.0] and 97.8% specificity 95% CI [95.5-99.1], whereas the assay targeting IgG1 showed the same sensitivity (100% 95% CI [76.8-100.0]) and a slightly higher specificity (99.1% 95% CI [97.3-99.8]). The other IgG subclasses (IgG2, IgG3, and IgG4) displayed a lower sensitivity and specificity. The sensitivity and specificity did not significantly differ between tests measuring total IgG and IgG1 (Exact McNemar's test; p > 0.05), with a concordance of 98.2%, represented by a Cohen's kappa of 0.83 (p < 0.001), indicating almost perfect agreement. Conclusion: ELISA targeting IgG1 can provide valuable information to clinicians in differentiating ALA from other parasitic diseases, cancers, cirrhosis, and viral hepatitis. However, enzyme-conjugated anti-human total IgG is cheaper than anti-human IgG subclasses. Therefore, we suggest that total IgG-based ELISA is sufficient for the routine serodiagnosis of human ALA and possibly other clinical manifestations of invasive amebiasis.


Subject(s)
Liver Abscess, Amebic , Humans , Liver Abscess, Amebic/diagnosis , Immunoglobulin G/analysis , Antibodies, Protozoan/analysis , Enzyme-Linked Immunosorbent Assay/methods , Serologic Tests/methods
16.
Trop Med Infect Dis ; 7(10)2022 Oct 17.
Article in English | MEDLINE | ID: mdl-36288049

ABSTRACT

Chronic human liver fluke infections caused by Opisthorchis viverrini and Clonorchis sinensis can last for decades and cause liver and biliary diseases, including life-threatening pathology prior to cholangiocarcinoma (CCA). CCA generally has a poor prognosis. Serological diagnosis can support parasitological examination in diagnosing disease and screening for the risk of CCA. Here, we present an improved and innovative lateral flow immunochromatographic test (ICT) kit that uses whole-blood samples (WBS) rather than serum to diagnose human opisthorchiasis, which also successfully diagnosed human clonorchiasis. This ICT includes a soluble worm extract of O. viverrini adults and colloidal-gold-labeled conjugates of the IgG antibody to evaluate the diagnostic values with simulated WBS (n = 347). Simulated WBS were obtained by the spiking infection sera with red blood cells. The diagnostic sensitivity, specificity, positive and negative predictive values, and accuracy for detecting opisthorchiasis were 95.5%, 87.0%, 80.5%, 97.2%, and 90.1%, respectively. For clonorchiasis, these findings were 85.7%, 87.0%, 53.6%, 97.2%, and 86.8%, respectively. Combined for both diseases, they were 93.2%, 87.0%, 84.0%, 94.6%, and 89.6%, respectively. The ICT kit can possibly replace the ICT platforms for antibody detection in serum samples in field surveys in remote areas where sophisticated equipment is not available.

17.
Orphanet J Rare Dis ; 17(1): 393, 2022 10 27.
Article in English | MEDLINE | ID: mdl-36303188

ABSTRACT

BACKGROUND: Eosinophilic meningitis (EOM) is a rare neurological disease that can be misdiagnosed or underdiagnosed. Based on reported cases in the literature, there have been 2,827 cases worldwide since 1945. There are limited data on the prevalence and trends of EOM in a real-world setting, even in Thailand, the country with the highest prevalence of EOM. Therefore, this study aimed to evaluate the prevalence of EOM and EOM epidemiological data in a real-world setting. METHODS: This was a pragmatic, retrospective analytical study using a national database. We retrieved EOM epidemiological data reported from government hospitals to Thailand's Bureau of Epidemiology, within the Ministry of Public Health's Department of Disease Control (DDC), between 2014 and 2019. The study was conducted by retrieving the data of all patients diagnosed with EOM and reported to the DDC. Diagnosis of EOM is made clinically by evidence of eosinophils of 10% or more of the total white blood cells in cerebrospinal fluid. Details of each patient were retrieved from the 506 Report Form, including age, month of reported case, zone of country, occupation, and mortality. Data regarding infection rate in each year and each zone were reported in rate/100,000 population, while data regarding age, month of reported case, and occupation were reported by year. Differences between means of age group, month of reported case, and occupation were tested by one-way analysis of variance (ANOVA). For those factors with significant differences among groups, Bonferroni method was used to compute pairwise differences. RESULTS: There were 1,083 EOM cases reported in Thailand during the six-year study period. The average annual incidence of EOM was 180.5 cases, or 0.27 cases/100,000 population. The northeast zone had the highest rate, with 0.89/100,000 population. The common age groups were 25-54 years, with the highest rate among the 35-44 age group, with a mean of 38.3 persons/year. These age groups were significantly different from other age groups (F value 39.23; p < 0.001). A relatively high cumulative monthly incidence (> 100 cases) was seen in four months, including January (117 cases), September (103 cases), October (112 cases), and November (103 cases), though these rates were not significantly different from the other months' rates. Regarding occupation, the top two occupations with EOM diagnoses were farmers and laborers, which were significantly different from other occupations (F value 99.95; p < 0.001). There was no reported case of death during the study period. CONCLUSION: EOM is common in Northeast Thailand among people of working age. The disease can be found throughout the year but is more common in the last quarter of the year. Farmers and laborers have the highest infection rate. To better understand the burden and outcomes of EOM, a national EOM reporting system with a better reporting form is required in endemic countries. Such a report form should include more details on risk exposure, symptoms, signs, treatment, and outcomes.


Subject(s)
Meningitis , Humans , Adult , Middle Aged , Retrospective Studies , Thailand/epidemiology , Meningitis/epidemiology , Incidence , Databases, Factual
18.
Bioconjug Chem ; 33(11): 2103-2112, 2022 11 16.
Article in English | MEDLINE | ID: mdl-36273419

ABSTRACT

In this work, the covalently oriented conjugation of monoclonal Listeria monocytogenes antibody (mAb-Lis) to amino-terminated oligo(ethylene glycol)-capped gold nanoparticles (NH2-TEG-AuNPs) was studied. After NH2-TEG-AuNPs were synthesized, the amino-terminated ligands on the particles were then linked to the carboxyl groups in the mAb-Lis through EDC/NHS chemistry. By maintaining the pH of the solution at ∼5, the Fc region of the antibody could preferably attach to the particle surface, providing a specific Fab region that was available for binding with the target pathogen. The resulting mAb-NH-TEG-AuNPs could act as a colorimetric probe for L. monocytogenes based on a particular antigen-antibody interaction, which resulted in a drastic aggregation of particles. This caused the color of the colloidal solution to transition from red-pink to purple or even gray depending on the pathogen concentration. To perform quantitative analysis, the absorbance ratio of A650/A534 was monitored as a function of L. monocytogenes concentration using a spectrophotometer. The detection limit was very low at 11 CFU/mL. Furthermore, a lateral flow strip (LFS) was fabricated as a portable device for onsite utilization. LFS detection could be completed by the naked eye and by a smartphone. The detection limit of LFS was estimated to be 103 CFU/mL. Our methods exhibited a substantial improvement in sensitivity compared to that of previous studies on immuno-based nanoparticles. The assay could be completed in 15 min, and no cross reactivity by any pathogen was found. Hence, the designed AuNPs exhibit great promise for use in monitoring L. monocytogenes for food safety and in other applications.


Subject(s)
Listeria monocytogenes , Metal Nanoparticles , Gold , Colorimetry/methods , Antibodies
19.
Parasite Epidemiol Control ; 19: e00272, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36133000

ABSTRACT

Introduction: Eosinophilic meningitis (EOM) is an emerging infectious disease worldwide. The most common cause of EOM is infection with Angiostrongylus cantonensis One possible method of monitoring and control of this infection is surveillance and prediction. There are limited data on national surveillance and predictive models on EOM. This study aimed to develop an online surveillance with a predictive model for EOM by using the national database. Methods: We retrospectively retrieved reported cases of EOM from all provinces in Thailand and quantified them by month and year. Data were retrieved from Ministry of Public Health database. We developed a website application to explore the EOM cases in Thailand including regions and provinces using box plots. The website also provided the Autoregressive Integrated Moving Average (ARIMA) models and Seasonal ARIMA (SARIMA) models for predicting the disease cases from nation, region, and province levels. The suitable models were considered by minimum Akaike Information Criterion (AIC). The appropriate SARIMA model was used to predict the number of EOM cases. Results: From 2003 to 2021, 3330 EOM cases were diagnosed and registered in the national database, with a peak in 2003 (median of 22 cases). We determined SARIMA(1,1,2)(2,0,0)[12] to be the most appropriate model, as it yielded the fitted values that were closest to the actual data. A predictive surveillance website was published on http://202.28.75.8/sample-apps/NationalEOM/. Conclusions: We determined that web application can be used for monitoring and exploring the trend of EOM patients in Thailand. The predictive values matched the actual monthly numbers of EOM cases indicating a good fit of the predictive model.

20.
Am J Trop Med Hyg ; 107(2): 370-372, 2022 08 17.
Article in English | MEDLINE | ID: mdl-35895399

ABSTRACT

Intestinal capillariasis is an emerging fish-borne helminthic disease caused by the round worm Capillaria philippinensis. Chronic infection may lead to death if the disease is misdiagnosed and inappropriate treatment is given. We used a rapid lateral-flow immunochromatographic test for screening of intestinal capillariasis in patients with chronic diarrhea. We screened 292 chronic diarrhea patients who had visited hospitals in Thailand. Sixty-six (22.6%) cases were positive according to the kit. All positive patients received mebendazole at 200 mg twice per day for 30 consecutive days or albendazole at 200 mg twice per day for 10 consecutive days. Later, stool concentration techniques, used to examine stool samples from all serologically positive individuals on three consecutive days, revealed C. philippinensis eggs, larvae, and/or adults. The kit is useful for screening and rapid diagnosis of intestinal capillariasis in chronic diarrhea patients in an endemic area for prevention of serious disease and facilitates treatment.


Subject(s)
Enoplida Infections , Intestinal Diseases, Parasitic , Animals , Intestinal Diseases, Parasitic/diagnosis , Intestinal Diseases, Parasitic/epidemiology , Thailand/epidemiology , Prevalence , Diarrhea/epidemiology , Immunoassay , Enoplida Infections/diagnosis , Enoplida Infections/drug therapy , Enoplida Infections/epidemiology
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