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Infections by the basidiomycete yeast Cryptococcus neoformans are life-threatening diseases claiming more than 600,000 lives every year. The most common manifestation is cryptococcal meningitis in AIDS patients. Diagnosis primarily relies on antigen testing from serum and cerebrospinal fluid (CSF). Current guidelines recommend rapid antigen testing with a focus on point-of-care assays. Over the recent years, a range of new lateral flow assays (LFAs) was launched. There is still a lack of data evaluating the CE-certified Biosynex RDT CryptoPS LFA. We compared the performance of this LFA with a latex agglutination assay (LAA; Latex-Cryptococcus Antigen Detection System, IMMY) from blood and CSF samples. Blood and/or CSF samples of 27 patients with proven cryptococcal infections caused by different species and blood–CSF pairs of 20 controls were tested applying LFA and LAA. Upon combined analysis of blood and CSF, both assays were able to identify all C. neoformans infections. Based on CSF analysis only, the LFA and the LAA had sensitivities of 100% and 93%. Neither test gave false-positive results nor was reactive in two cases of C. non-neoformans/non-gattii species infections. Both assays have high sensitivities and specificities for the diagnosis of C. neoformans infection. Contrarily to the IMMY LAA, the RDT CryptoPS LFA is suitable as a point-of-care test but is limited in the quantification of antigen reactivity.

Background Lactoferrin (LF) in human milk has various biological properties and contributes to the prevention of preterm birth complications. Enzyme-linked immunosorbent assay (ELISA) is one of the most commonly used methods to measure LF in human milk, but this method is time-consuming and laborious. In Japanese human milk banks, the concentration of LF in donor human milk (DHM) is measured routinely. Here, we reported a rapid, simple, and accurate method for determining LF in human milk using a new reagent based on a latex agglutination assay. Methods We obtained 208 human milk pools from 148 mothers, and samples were collected before and after Holder pasteurization. Milk samples were diluted 100- or 200-fold and LF concentrations were measured by a latex agglutination assay using an automated analyzer. The reagent was validated in terms of repeatability, linearity, detection limit, recovery, and comparison with ELISA. Results The coefficient of variation (CV) for intra-assay precision ranged from 0.6 to 5.0% in human milk with high, medium, and low LF concentrations. The linearity was also tested by serial sample dilution and was confirmed up to 16 µg/mL with a detection limit of 0.2 µg/mL. The recovery rates in a spiked recovery test were ranged from 90 to 120% at high, medium, and low concentrations of LF. Furthermore, a strong correlation was observed between LF levels determined by the latex agglutination assay and ELISA ( r  = 0.978, p  < 0.001, n  = 255). The regression equation was y = 0.991x + 0.545 (r 2  = 0.974, p  < 0.001). Compared with ELISA, the latex agglutination assay reduces the measurement time by 160 min and the cost by 55%. Conclusions The latex agglutination assay used to determine LF in human milk is rapid, simple, and accurate enough to be used routinely. Its use may contribute to the quick and easy provision of appropriate DHM to preterm infants.

The monoclonal antibody-based latex agglutination tests targeting a high molecular weight exopolysaccharide antigen of Burkholderia pseudomallei are commercially available. The tests are primarily used in routine diagnosis of melioidosis in major hospitals in Thailand and some endemic countries. Being a rapid test, this technique was employed as a presumptive test to identify colonies of B. pseudomallei among many others grown from soil specimens collected from southern Thailand. Cross-reactivity of these tests with other soil bacteria was a concern since it complicated the identification of B. pseudomallei. Here, we describe the cross-reactivity of two commercial latex agglutination tests for melioidosis with B. territorii, B. pseudomultivorans, B. multivorans and B. cenocepacia isolates from soil.

•Introduction led to in a rapid reduction of H. influenzae type b mengintis.•Impact was larger than expected from vaccine coverage, suggesting herd immunity.•Introduction resulted in reduction of probable meningitis with no etiology found. Haemophilus influenzae type b was the leading cause of bacterial meningitis in infants and children below the age of two years prior to the introduction of H. influenzae type b conjugate vaccines. In December 2011, the Indian government introduced H. influenzae b vaccine in the state of Tamilnadu. A prospective surveillance for bacterial meningitis was established at the Institute of Child Health in Chennai to evaluate the etiology of meningitis and impact of the vaccine. Infants aged one to 23 months who were admitted to the hospital with symptoms of suspected bacterial meningitis were enrolled and lumbar puncture was performed. Cerebrospinal fluid samples were analyzed for white blood cells, protein, and glucose. Bacterial culture and a latex agglutination test for common bacterial pathogens were performed. Between January 2009 and March 2014, 4,770 children with suspected bacterial meningitis were enrolled. Prior to the introduction of the vaccine, an average of 11.7 cases of H. influenzae b meningitis and 31.1 cases of probable meningitis with no etiology were identified each year. After introduction, the number of cases were reduced by 79% and 44% respectively. The average H. influenzae b vaccine coverage after introduction was 69% among all children with clinically suspected meningitis. In contrast, the mean number of aseptic meningitis and pneumococcal meningitis cases remained stable throughout the pre and post vaccination period; 28.2 and 4.8 per year, respectively. H. influenzae b conjugate vaccine reduced the number of cases of H. influenzae b meningitis and probable meningitis within the first two years of its introduction. The impact against meningitis was higher than the vaccination rate, indicating indirect effects of the vaccine. India has recently scaled up the use of Hib conjugate vaccine throughout the country which should substantially reduce childhood meningitis rates further in the country.

Background Biomolecules in body fluids such as gingival crevicular fluid and saliva are used for the diagnosis of periodontitis. Saliva is easy to collect and salivary biomarkers are useful in screening periodontitis. The suitable salivary biomarkers and their measuring system are important for screening periodontitis in mass dental examination. Therefore, this study examined the potential of few salivary biomarkers for screening and diagnosis of periodontitis and aimed to evaluate more effective biomarkers and their measuring methods in dental examination. Methods Ninety-three individuals with and without periodontitis participated in this clinical examination and were classified into the non-periodontal diseases or stage I periodontitis (control, n  = 26) and stage II-IV periodontitis groups (periodontitis, n  = 67) after periodontal examinations. Unstimulated saliva samples were collected from participants. The levels of salivary biomarkers including calprotectin, hemoglobin (Hb), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), alanine aminotransferase and aspartate aminotransferase (AST) were automatically measured using the latex agglutination turbidimetric immunoassay (LATIA) and enzyme assay systems. Differences in clinical indicator and biomarker levels in the control and periodontitis groups and their correlations were statistically analyzed. A receiver operating characteristic (ROC) analysis of the ability of salivary biomarkers to predict periodontitis was also performed. Results Salivary calprotectin, Hb, LDH, ALP and AST levels were significantly higher in the periodontitis group than that in the control group. At the initial stage of periodontitis, a significant difference was only observed in calprotectin levels. Calprotectin and LDH levels strongly correlated with clinical indicators including probing pocket depth, clinical attachment level, bleeding on probing, gingival index and periodontal inflamed surface area with high correlation coefficient (calprotectin: 0.582–0.660, LDH: 0.534–0.614). Calprotectin showed a higher area under the ROC curve value (0.894), with 91% sensitivity and 73% specificity, than the other salivary biomarkers. Conclusions Salivary calprotectin showed a high effectiveness for the diagnosis of periodontitis, and the measurement of salivary calprotectin using the LATIA system that is a high throughput method is suitable for population-based screening of periodontal diseases.

Background Toxoplasmosis in camels is an important zoonotic infection with considerable economic and public health impacts, particularly in the pastoral regions of Ethiopia. Methods A cross-sectional study was conducted in selected districts of the Borana zone, Southern Oromia of Ethiopia, to estimate the Toxoplasma gondii seroprevalence infection and associated risk factors in camels. Accordingly, 352 camel blood samples were randomly collected. Then, the sera were separated from the blood and analyzed using a latex agglutination test. Results According to this study, the seroprevalence of camel T. gondii in the study districts using the latex agglutination test was 7.9%. The current study revealed that the T. gondii seroprevalence was relatively higher in the Miyo (9.1%) district. The males (8.9%), the adult age group (5.8%), and the poor-condition camels (21.0%) had higher seroprevalence. There was a statistically significant association between the body condition and the seroprevalence of camel T. gondii ( p  = 0.016). However, no statistically significant difference was noted between the seroprevalence of camel T. gondii and the peasant association, districts, age, and sex. Conclusion This study revealed a significant prevalence of Toxoplasma gondii infection in camels, posing a potential public health risk in the study area. To effectively control the disease and minimize its impact on both animal and human health, it is important to raise public awareness, implement practical biosecurity measures, and conduct further comprehensive studies.

Background Melioidosis is a serious bacterial infection caused by Burkholderia pseudomallei , a gram-negative bacterium commonly found in soil and water. It can affect both humans and animals, and is endemic in regions such as Southeast Asia and Northern Australia. In recent years, there have been reports of an emergence of human melioidosis in other areas, including New Caledonia. Results During standard laboratory analysis in New Caledonia in 2021, a strain of B. pseudomallei was isolated from a goat. The strain was characterized using both MLST and WGS techniques and was found to cluster with previously described local human strains from the area. In parallel, several serological tests (CFT, ELISA, Luminex (Hcp1, GroEL, BPSS1840), arrays assay and a latex agglutination test) were performed on animals from the farm where the goat originated, and/or from three other neighboring farms. Using two commercial ELISA kits, seropositive animals were found only on the farm where the infected goat originated and tests based on recombinant proteins confirmed the usefulness of the Hcp1 protein for the diagnosis of melioidosis in animals. Conclusions Despite the regular reports of human cases, this is the first confirmed case of melioidosis in an animal in New Caledonia. These results confirm the presence of the bacterium in the region and highlight the importance of vigilance for both animal and human health. It is critical that all health partners, including breeders, veterinarians, and biologists, work together to monitor and prevent the spread of the disease.

The present study is focused to express the recombinant Leptospira surface adhesion-44 (rLsa44) protein of pathogenic Leptospira interrogans serovar Pomona in Escherichia coli and its diagnostic application in the latex agglutination test (LAT) system to detect anti-leptospiral antibodies in the bovine sera. The Lsa44 protein-coding gene sequences were amplified, the purified amplicon was initially cloned into the pGEM-T Easy vector, and subsequently to the pETite vector for the expression of rLsa44 in Escherichia coli . After induction with 1 mM isopropyl-β-D-thiogalactoside (IPTG), the expressed recombinant protein with a molecular weight of ~42 kDa was purified by the Ni-NTA affinity chromatography, characterized by SDS-PAGE and confirmed by Western blot using Leptospira specific sera. The sensitized latex beads (0.8 µm) coated with rLsa44 protein were prepared and assessed for their suitability as a diagnostic antigen in the LAT. Ten microliters of sensitized beads were used with test serum for agglutination, the results were recorded for 3 min and positive results were rated as +++, ++, +, if the clumps appeared after 1, 2, and 3 min, respectively. The rLsa44-LAT was evaluated for detection of anti-leptospiral antibodies using known Microscopic Agglutination Test (MAT) with positive ( n = 74) and negative ( n = 91) samples from apparently healthy animals, which revealed the relative diagnostic sensitivity (DSn) of 88% and specificity (DSp) of 84%. Furthermore, on the evaluation of standardized rLsa44-LAT using MAT-tested sera (positive-62 and negative-64) from cattle associated with a history of reproductive disorder, a DSn of 94% and DSp of 91% were observed. The developed rLsa44-LAT is a simple and rapid test that can be applied as a diagnostic tool at the field level as an on-site test after its extensive evaluation. This first study describing the expressed rLsa44 protein in Escherichia coli and used as a diagnostic antigen in LAT for the detection of anti-leptospiral antibodies in the sera of bovine as a preliminary screening test for leptospirosis.

Human trichinellosis is a worldwide foodborne public health threat. Detecting circulating antigens of Trichinella spiralis “ T. spiralis ” allows for an early diagnosis before larval encystation develops in skeletal muscles. For the first time, the present study aimed to formulate an effective nanomagnetic beads based-ELISA and -latex agglutination test (NMB-ELISA and NMB-LAT) to recognize T. spiralis adult worm crude extract antigen (AWCEA) in sera of experimentally infected mice. The study included thirty-eight mice classified into 3 groups; T. spiralis -infected group (GI) which was euthanized 6, 8, 10, 12, 14 days post-infection (dpi), other parasitic infections group (GII) and healthy control group (GIII). Rabbit anti- T. spiralis polyclonal antibodies (pAbs) were utilized to detect AWCEA in serum samples by sandwich ELISA, NMB-ELISA, and NMB-LAT. Using NMB - ELISA, AWCEA was detected in sera collected at 6 and 8 dpi, with a sensitivity of 50% and 75%, respectively, and a specificity of 100%. Whereas, sandwich ELISA and NMB - LAT couldn’t detect the antigen at the same time intervals. Both ELISA formats were able to detect the antigen in samples collected at 10, 12, and 14 dpi with a sensitivity of 100% for NMB-ELISA and 25%, 75%, and 100% respectively, for sandwich-ELISA. Yet, NMB-LAT couldn't detect AWCEA until 12 dpi with a sensitivity of 50% and specificity of 75%. In conclusion, NMB-ELISA is a promising sensitive tool for early and specific diagnosis of acute trichinellosis. The use of NMB-LAT could be a helpful screening procedure in field surveys.

Toxoplasma gondii is a major neglected parasitic infection occurring in settings of extreme poverty in Africa. Apart from causing reproductive failure in animals it is also a significant zoonotic concern. The objective of this study was to determine the seroprevalence and associated risk factors of T. gondii infection in cats, chickens, goats, sheep and pigs in the southeast of South Africa, of which little is known. Sera was obtained from 601 domestic animals including 109 cats, 137 chickens, 128 goats, 121 sheep and 106 pigs managed under different production systems in different agro-ecological regions and evaluated by the Toxoreagent, a latex agglutination test for T. gondii antibody detection. Household-level and animal-level data were collected by interviewing animal owners and/or herders using a closed-ended questionnaire. The study revealed an overall farm seroprevalence of 83.33% (125/150 farms) with the highest rate of infection for the parasite found in sheep with 64.46% (78/121), followed by goats with 53.91% (69/128), pigs with 33.96% (36/106), cats with 32.11% (35/109 cats) and chickens with 33.58% (46/137). The risk factors that were found to be statistically significant (p 0.05) to different species of seropositivites were age, location, climate, animal production system, rodent control, seropositive cat, cat-feed access and cat faecal disposal. The relatively high seroprevalence of T. gondii detected in this region suggests that domestic animals may pose a substantial public health risk through the consumption of T. gondii-infected raw meat as well as via contact with cat faeces.