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Veterinary Research Communications Dec 2023The microscopic agglutination test (MAT) assay is adopted as a world-wide reference test for the serodiagnosis of leptospirosis in humans and animals. One of the main...
The microscopic agglutination test (MAT) assay is adopted as a world-wide reference test for the serodiagnosis of leptospirosis in humans and animals. One of the main limitations of MAT is the lack of sensitivity and serodiagnostic antigens should be periodically updated with locally circulating serovars in order to optimise its performance. The aim of this study was to determine the need to implement the antigen panel currently adopted in Northern Italy for the diagnosis of Leptospira infection in dogs. For this purpose, a group of 288 dogs with and without clinical signs potentially consistent with Leptospira infection or found to have an increased C-reactive protein (CRP) serum concentration, sampled in 2013-2016 in Northern Italy, were tested by MAT comparing the results obtained with a nine antigens panel (Australis-Bratislava, Ballum-Ballum, Canicola-Canicola, Grippotyphosa-Grippotyphosa, Icterohaemorrhagiae-Copenhageni, Icterohaemorrhagiae-Icterohaemorrhagiae, Sejroe-Hardjo, Pomona-Pomona and Tarassovi-Tarassovi serovars) routinely adopted and a panel expanded to 27 antigens. In general, the antigen panel currently adopted in Northern Italy for the routine MAT assay resulted adequate for the diagnosis of Leptospira infection in dogs. The main exception concerns the Sejroe serogroup, with the Saxkoebing and Sejroe serovars that were more effective than Hardjo for diagnosis in dogs and whose inclusion in the antigen panel is recommended. Among other antigens evaluated in this study, Cynopteri serovar was detected with high frequency but its pathogenic role in dogs and as public health threat deserve further investigation.
Topics: Humans; Animals; Dogs; Serogroup; Antibodies, Bacterial; Leptospirosis; Agglutination Tests; Serologic Tests; Dog Diseases
PubMed: 36929155
DOI: 10.1007/s11259-023-10103-3 -
A Quantitative Human Red Blood Cell Agglutination Assay for Characterisation of Galectin Inhibitors.International Journal of Molecular... Jun 2024Galectins are a family of beta-galactoside-binding proteins that are characterised by their carbohydrate recognition domain (CRD) and include galectin-1 and galectin-3....
Galectins are a family of beta-galactoside-binding proteins that are characterised by their carbohydrate recognition domain (CRD) and include galectin-1 and galectin-3. These galectins have been implicated in numerous diseases due to their pleiotropic nature, including cancer and fibrosis, with therapeutic inhibitors being clinically developed to block the CRD. One of the early methods developed to characterise these galectins was the hemagglutination of red blood cells. Although it is insightful, this approach has been hampered by a lack of sensitivity and accurate quantification of the agglutination observed. In this study, we aimed to validate a more precise and quantitative method to enable the further investigation of differences between galectins in respect to agglutination induction in different blood groups, as well as the characterisation of small molecule inhibitors. Quantification of hemagglutination was shown to be optimal using U-bottom plates imaged and analysed with FIJI ImageJ rather than flat-bottom plates read for absorbance on an optical density plate reader. Galectin-3-induced red blood cell agglutination efficacy increased significantly from blood group O to A to B. However, for both the galectin-1 monomer and concatemer, a more comparable effect was observed between blood group B and O, but with more potent effects than in blood group A. Inhibition assays for both galectin-3 and galectin-1 induced-hemagglutination were able to demonstrate clear concentration responses and expected selectivity profiles for a set of small-molecule glycomimetics, confirming the historical profiles obtained in biochemical binding and functional cellular assays.
Topics: Humans; Erythrocytes; Hemagglutination; Galectins; Galectin 1; Galectin 3; Agglutination Tests; Hemagglutination Tests; Agglutination
PubMed: 38928462
DOI: 10.3390/ijms25126756 -
Scientific Reports Jul 2023Thailand is known to be endemic for leptospirosis. This bacterium may pose a potential risk to transfusion safety. This study was a cross-sectional study examining the...
Thailand is known to be endemic for leptospirosis. This bacterium may pose a potential risk to transfusion safety. This study was a cross-sectional study examining the seroprevalence of leptospirosis among Thai blood donors. A total of 1053 serum specimens collected from blood donors residing in 5 regions of Thailand during March to September 2020 were included in this study. All samples were tested for the presence of antibodies to 22 leptospiral serovars using the microscopic agglutination test (MAT) and anti-Leptospira IgG antibodies using commercially available enzyme immunoassay. We found no evidence of recent exposure to Leptospira spp. in sera of healthy Thai blood donors by MAT, including those in higher-risk areas. However, in this same group, we did find small numbers of past exposure (1.7%) to Leptospira spp. by IgG ELISA. According to the findings of this study, there is currently no evidence for implementing new blood banking procedures to identify possible carriers in Thailand, however these should be continually monitored and revised according to the infectious disease burden in each country. It should be noted that there was a difference in the occupation rate between the general population reported in Thailand and blood donors in this study; it may not reflect the actual situation in the country.
Topics: Humans; Seroepidemiologic Studies; Blood Donors; Cross-Sectional Studies; Leptospirosis; Leptospira; Antibodies, Bacterial
PubMed: 37524788
DOI: 10.1038/s41598-023-39461-3 -
Clinical Microbiology and Infection :... Dec 2023Serological tests for syphilis detect mainly total Ig, IgM or IgG antibodies. We aimed to evaluate the specific IgA response in syphilis patients according to disease...
OBJECTIVES
Serological tests for syphilis detect mainly total Ig, IgM or IgG antibodies. We aimed to evaluate the specific IgA response in syphilis patients according to disease stage.
METHODS
A serum IgA-enzyme immunoassay was developed using commercially available microplates coated with recombinant treponemal antigens and an anti-IgA-conjugate. To define a cut-off, we used 91 syphilis positive and 136 negative sera previously defined by the rapid plasma reagin and the Treponema pallidum particle agglutination results. Then we determined the intra- and inter-assay precisions, diagnostic sensitivity according to the clinical stage (in 66, 55 and 42 sera from primary, secondary and latent syphilis patients, respectively) and specificity (in 211 sera from people with conditions different to syphilis). IgA values were further measured in 71 sera from patients with previously treated syphilis.
RESULTS
The newly developed IgA-enzyme immunoassay showed a good discrimination between negative and positive samples with intra- and inter-assay variation coefficients <20%. The sensitivity was 80.3% (95% CI, 70.0-90.6), 100.0% (95% CI, 99.1-100.0) and 95.2% (95% CI, 87.6-100.0) in primary, secondary and latent syphilis, respectively, and the specificity was 98.1% (95% CI, 96.0-100.0). Further, IgA values were negative in 61.3% (38/62) of patients with previously treated syphilis.
DISCUSSION
Our findings suggest serum IgA as a sensitive and specific marker of syphilis and its detection could be used as a screening assay for active infection. Further evaluation is needed in prospective longitudinal field studies.
Topics: Humans; Treponema pallidum; Syphilis; Prospective Studies; Syphilis Serodiagnosis; Immunoglobulin A; Antibodies, Bacterial; Sensitivity and Specificity
PubMed: 37611864
DOI: 10.1016/j.cmi.2023.08.015 -
International Ophthalmology Dec 2023The purpose of the study was to evaluate the incidence of ocular syphilis as well as diagnostic parameters, comorbidities, and visual outcomes over a 10-year time period...
PURPOSE
The purpose of the study was to evaluate the incidence of ocular syphilis as well as diagnostic parameters, comorbidities, and visual outcomes over a 10-year time period in West Virginia.
METHODS
A retrospective chart review included 25 eyes of 17 patients with ocular syphilis between 2010 and 2020.
RESULTS
The incidence of systemic syphilis at a large tertiary referral center has increased from 27 cases in 2010 to 105 cases in 2020. Seventeen patients were identified with ocular syphilis. Bilaterality was present in 47.1% of cases. In this study, 70.6% of patients were male and 29.4% were female. The median age of presentation was 40.2 years (range 21-63). Panuveitis was the most common (60.0%) followed by isolated anterior uveitis (16.0%), chorioretinitis (12.0%), inner retinitis (4.0%), and papillitis (8.0%). Forty percent of patients had visual acuity worse than 20/400 on presentation. Post-treatment visual acuity improved in all patients. Rapid plasma reagin (RPR) and Treponema pallidum particle agglutination (TP-PA) tests were positive in 84.6% and 100% of cases, respectively. CSF venereal disease research laboratory (VDRL) was positive in 36.4%, CSF pleocytosis was present in 72.7%, and elevated CSF protein was observed in 81.8%. Human immunodeficiency virus (HIV) co-infection was present in 31.3%. A majority of patients experienced maculopapular rash and/or history of genital chancre. The anatomic classification of presenting uveitis (anterior, intermediate, posterior, and panuveitis) did not correlate with clinical variables including age, gender, HIV status, serologic test, presence of rash, or year of diagnosis (p > 0.05).
CONCLUSION
Ocular syphilis is becoming increasingly prevalent and can present with a variety of ocular findings; therefore, it should be considered in the differential diagnosis for patients with ocular inflammation. Visual prognosis is excellent with timely diagnosis and treatment.
Topics: Humans; Male; Female; Young Adult; Adult; Middle Aged; Syphilis; Retrospective Studies; Chorioretinitis; Panuveitis; Endophthalmitis; HIV Infections; Exanthema
PubMed: 37773472
DOI: 10.1007/s10792-023-02759-2 -
Transfusion Medicine (Oxford, England) Oct 2023Pre-transfusion testing (PTT) encompasses a set of mandatory laboratory tests performed before red blood cell transfusion. The antibody screen, one component of PTT,...
BACKGROUND
Pre-transfusion testing (PTT) encompasses a set of mandatory laboratory tests performed before red blood cell transfusion. The antibody screen, one component of PTT, commonly includes a 10-20 min incubation. The primary aim of this study was to determine if this period can be reduced when using current immunohematology methodologies.
METHODS AND MATERIALS
Antibody screens were performed on reagent samples using Glass or Gel-based column agglutination technologies (CAT) and a solid phase red cell adherence (SPRCA) assay, with incubation periods of 1, 5, 10 and 15 min, and 20 min (SPRCA assay only). For each method, the shortest period producing a minimum of a 1+ reaction with all reagent samples was considered optimal. The sensitivity of each assay using the optimal period was calculated after performing antibody screens on 100 patient samples.
RESULTS AND DISCUSSION
It was demonstrated that the incubation period in the SPRCA and Glass CAT systems can be reduced to 5 and 10 min, respectively, while achieving high assay sensitivity (98.9% in both). The incubation period in the Gel CAT system cannot be reduced from 15 min. Significant association between titre and reaction strength was observed for all three screening methods (p < 0.001 for both CAT methods, p = 0.041 for SPRCA). This study demonstrates that the incubation period used in the antibody screen can be reduced when using systems employing the Glass CAT and SPRCA methods, without affecting assay sensitivity. If confirmed, it could result in faster completion of PTT.
Topics: Humans; Blood Grouping and Crossmatching; Erythrocytes; Time Factors; Sensitivity and Specificity; Antibodies
PubMed: 37728214
DOI: 10.1111/tme.13009 -
Acta Tropica Dec 2023Chlamydiosis and brucellosis induced abortions have resulted in significant economic losses in the global livestock industry. Although there have been numerous reports...
Chlamydiosis and brucellosis induced abortions have resulted in significant economic losses in the global livestock industry. Although there have been numerous reports on these two diseases in ruminants in China, limited information is available regarding the prevalence of Chlamydia abortus (C. abortus) and Brucella spp. infection in pigs. This study aimed to investigate the prevalence of C. abortus and Brucella spp. infections in pig serum using serology and to identify potential risk factors. In total, 2816 serum samples were collected from 12 provinces in China. The presence of C. abortus antibodies was determined using an enzyme-linked immunosorbent assay (ELISA), while the presence of Brucella spp. antibodies was examined using the Rose Bengal Plate Test (RBPT) and the Standard Agglutination Test (SAT). The seroprevalences of C. abortus and Brucella spp. were 8.38 % (236/2816) and 0.11 % (3/2816), respectively. Geographical location, season, and age were found to be risk factors associated with C. abortus infection in pig herds in China (p<0.01), and the seropositive rate for C. abortus in sow herds was strongly associated with the occurrence of abortion (p<0.01). Overall, in China, pigs exhibit a higher seroprevalence of C. abortus, whereas the prevalence of Brucella is limited. This study represents the first comprehensive survey of C. abortus and Brucella spp. in pig herds in China that established potential risk factors and provided data for the prevention and control of intraspecies and interspecies transmission of C. abortus to humans.
Topics: Pregnancy; Humans; Swine; Animals; Female; Brucella; Seroepidemiologic Studies; Brucellosis; Risk Factors; Antibodies, Bacterial; China; Brucella abortus
PubMed: 37875168
DOI: 10.1016/j.actatropica.2023.107050 -
Journal of Food Protection Mar 2024Raw vegetables and herbs are exposure sources of foodborne pathogens. This study examined the prevalence and antimicrobial resistance of Escherichia coli in five types...
Raw vegetables and herbs are exposure sources of foodborne pathogens. This study examined the prevalence and antimicrobial resistance of Escherichia coli in five types of fresh vegetables and herbs: spearmint (Mentha spicata), leaf lettuce (Lactuca sativa var. crispa), coriander (Coriandrum sativum), Chinese cabbage (Brassica rapa subsp. pekinensis), and cucumber (Cucumis sativus). Produce samples (n = 300) were acquired from local open markets and supermarkets in Bangkok, Thailand. Each produce sample was preenriched in buffered peptone water and then enriched in E. coli broth. A loopful of the second enrichment was transferred onto selective media for subsequent confirmation and biochemical tests. The Kirby-Bauer disk diffusion test was employed to assess E. coli susceptibility to selected antibiotics (twelve antibiotics of seven classes). The latex agglutination test was performed to serotype the isolates for O157 and H7 antigens. A commercial test kit was used to determine the presence of Shiga toxin-producing E. coli (STEC). For all samples, the prevalence of E. coli was found to be 32.0% (96/300), whereby produce from supermarkets had a higher prevalence than those from local markets at 40.7% (61/150) and 23.3% (35/150), respectively. Among different types of produce, leaf lettuce had the highest E. coli prevalence at 36.7% (22/60), followed by cucumber and spearmint, coriander, and Chinese cabbage, at 35.0% (21/60), 35.0% (21/60), 35.0% (21/60), and 18.3% (11/60), respectively. Of the positive isolates, 27.1% (26/96) showed multidrug resistance. All isolates (100%) showed resistance to penicillin but varying resistant characteristics for tetracycline, ampicillin, and amoxicillin, with resistance rates of 31.3% (30/96), 31.3% (30/96), and 31.3% (30/96), respectively. Two of the 96 isolates (1.1%) were positive for the O157 antigen but negative for the H7 antigen. No STEC was observed. This study established baseline information regarding the prevalence of E. coli and its antimicrobial resistance profile in produce in Bangkok, Thailand.
Topics: Vegetables; Prevalence; Thailand; Escherichia coli O157; Anti-Bacterial Agents; Escherichia coli Infections; Shiga-Toxigenic Escherichia coli
PubMed: 38246524
DOI: 10.1016/j.jfp.2024.100229 -
Frontiers in Medicine 2023To prevent chronic brucellosis, this study analysed the changes in patient antibody titers, and the trajectories of biochemical indicators at different stages of...
OBJECTIVE
To prevent chronic brucellosis, this study analysed the changes in patient antibody titers, and the trajectories of biochemical indicators at different stages of brucellosis, identified relevant biomarkers, and explored risk factors affecting the prognosis of brucellosis patients.
METHODS
A prospective cohort study was conducted to follow 100 patients with acute brucellosis. Laboratory serological test results [taken with a serum (tube) agglutination test (SAT)] and biochemical parameters (liver function, renal function, and hematological system) were measured repeatedly at four-time points: 0 weeks-baseline survey, 6 weeks after the first treatment, 12 weeks after the second treatment, and 3 months after the third treatment. The changes in the antibody titres and biochemical parameters at each time point were analysed for trend changes.
RESULTS
One hundred patients with acute brucellosis were enrolled in this follow-up study, with 100% retention in follow-up. By the third follow-up, 21 patients had turned subacute and 11 had turned chronic. One-way repeated measures analysis of variance results showed statistically significant differences ( < 0.01) across the time points for the following five indicators: alanine aminotransferase, aspartate aminotransferase, total bilirubin, serum creatinine (SCr) and platelet count. The clinical symptoms of patients in the acute stage were mainly joint pain, fatigue, and fever, while those in the chronic stage complained primarily of joint pain and fatigue. The results of multivariate logistic analysis showed that joint pain [odds ratio (OR) = 3.652, 95% confidence interval (CI) =1.379-9.672], monoarticular pain (OR = 6.356, 95% CI = 4.660-8.669), elevated SCr (OR = 15.804, 95% CI = 1.644-151.966) and elevated haemoglobin (Hb) (OR = 1.219, 95% CI = 1.065-1.736) were risk factors for poor prognosis (not cured or chronic) in patients with brucellosis.
CONCLUSION
The trajectory of changes in patient SAT posirates and antibody titers can be used to distinguish patients with chronic brucellosis. The brucellosis is preventable and treatable, and the standard treatment can be effective in reducing the clinical symptoms of affected patients. If patients are not treated in a timely manner, joint pain, monoarticular pain, and elevated SCr are risk factors for patients who are not cured. Therefore, the treatment cycle for these patients should be extended.
PubMed: 38020155
DOI: 10.3389/fmed.2023.1110907 -
Future Microbiology 2024This study investigates the application of metagenomic next-generation sequencing (mNGS) in the diagnosis of neurobrucellosis (NB). We retrospectively analyzed...
This study investigates the application of metagenomic next-generation sequencing (mNGS) in the diagnosis of neurobrucellosis (NB). We retrospectively analyzed patients diagnosed with NB who underwent cerebrospinal fluid (CSF) mNGS testing in Xijing Hospital from 2015 to 2021. Among the 20 individuals included in the study, the serum rose bengal test was positive in 11 out of 16 cases, serum agglutination test was positive in 13 out of 16 cases, CSF culture was positive in 6 out of 11 cases, and CSF mNGS tests were positive in 18 out of 20 cases. CSF mNGS demonstrates superior sensitivity; therefore, it is recommended to collect CSF for mNGS testing prior to antibiotic therapy when NB is suspected.
Topics: Humans; High-Throughput Nucleotide Sequencing; Retrospective Studies; Metagenomics; Female; Male; Middle Aged; Adult; Brucellosis; Aged; Young Adult; Sensitivity and Specificity; Adolescent
PubMed: 38629898
DOI: 10.2217/fmb-2023-0177