[Use of LDBIO Diagnostics® TOXOPLASMA ICT IGG-IGM in association with Siemens Atellica® IM in toxoplasmosis serological screening]. / Place du test rapide LDBIO Diagnostics® TOXOPLASMA ICT IGG-IGM en complément de la sérologie Siemens Atellica® IM dans la stratégie diagnostique de la toxoplasmose.

According to French recommendations for serological screening of toxoplasmosis, some profiles must be confirmed by additional methods, extending the time taken to produce results. Thus, the Laborizon Bretagne technical platform in Nantes studied the place of the LDBIO Diagnostics® TOXOPLASMA ICT IGG-IGM (ICT) test in addition to Siemens Atellica® serology. IgG-/IgM+ and equivocal or weak positive IgG/IgM- (IgGEq/IgM-) profiles on Atellica® will be confirmed by ICT, Alinity® Abbott and Platelia® Biorad. Among the 66 IgGEq/IgM- profiles, the concordance is perfect between ICT and complementary techniques: 21 weak positives were confirmed positive, 8 equivocal were considered negative and 37 were confirmed positive. Concerning the 76 IgG-/IgM+ profiles, 68 are negative and 7 are positive by complementary techniques and ICT. One discordance was observed. The Atellica®/ICT combination allows excellent discrimination of IgG-/IgM+ and IgGEq/IgM serological profiles with consistent diagnostic orientation in 99.3% of cases. Only 1 sample was found to be discordant but required monitoring at 15 days. The observed performances are compatible with routine use. This test simplifies the analytical process, improves the time to obtain results, while guaranteeing an excellent level of quality.

Multicenter evaluation of the Toxoplasma gondii Real-TM (Sacace) kit performance for the molecular diagnosis of toxoplasmosis.

The molecular detection of Toxoplasma gondii DNA is a key tool for the diagnosis of disseminated and congenital toxoplasmosis. This multicentric study from the Molecular Biology Pole of the French National Reference Center for toxoplasmosis aimed to evaluate Toxoplasma gondii Real-TM PCR kit (Sacace). The study compared the analytical and clinical performances of this PCR assay with the reference PCRs used in proficient laboratories. PCR efficiencies varied from 90% to 112%; linearity zone extended over four log units (R2 > 0.99) and limit of detection varied from 0.01 to ≤1 Tg/mL depending on the center. Determined on 173 cryopreserved DNAs from a large range of clinical specimens, clinical sensitivity was 100% [106/106; 95 confidence interval (CI): 96.5%-100%] and specificity was 100% (67/67; 95 CI: 94.6%-100%). The study revealed two potential limitations of the Sacace PCR assay: the first was the inconsistency of the internal control (IC) when added to the PCR mixture. This point was not found under routine conditions when the IC was added during the extraction step. The second is a lack of practicality, as the mixture is distributed over several vials, requiring numerous pipetting operations. Overall, this study provides useful information for the molecular diagnosis of toxoplasmosis; the analytical and clinical performances of the Sacace PCR kit were satisfactory, the kit having sensitivity and specificity similar to those of expert center methods and being able to detect low parasite loads, at levels where multiplicative analysis gives inconsistently positive results. Finally, the study recommends multiplicative analysis in particular for amniotic fluids, aqueous humor, and other single specimens.

A systematic review and meta-analysis of the validation of serological methods for detecting anti-Toxoplasma gondii antibodies in humans and animals.

Toxoplasma gondii is a paradigmatic zoonotic parasite from the One Health perspective, since it is broadly distributed and virtually infects all warm-blooded species. A wide variety of serological techniques have been developed to detect T. gondii infection in humans and animals. Our aim was to describe and compare the main characteristics of these serological tests and validation processes and to critically analyze whether these tests meet the standards required to ensure an accurate serological diagnosis. The current systematic review and meta-analysis included 134 studies that were published from 2013 to 2023. QUADAS 2 tool was used to evaluate the quality of the included studies. A total of 52 variables related to the characteristics of the techniques and analytical and diagnostic validation parameters were studied. A wider panel of tests was developed for humans, including techniques exclusively developed for humans that involve costly equipment and the measurement of different Ig isotypes that are considered biomarkers of congenital toxoplasmosis. Studies conducted in humans frequently employed commercial techniques as reference tests, measured different immunoglobulin isotypes with a predominance for IgG (>50%) and discriminated between acute and chronic infections. In animals, the most commonly used reference techniques were in-house tests, which almost exclusively detected IgG. Common limitations identified in a large number of studies were some misunderstandings of the terms "gold standard" and "reference test" and the absence of information about the negative and positive control sera used or the exact cutoff employed, which were independent of the quality of the study. There is a lack of analytical validation, with few evaluations of cross-reactivity with other pathogens. Diagnostic odds ratio values showed that indirect ELISA based on native or chimeric antigens performed better than other tests. The reproducibility of serological test results in both humans and animals is not guaranteed due to a lack of relevant information and analytical validation. Thus, several key issues should be considered in the future, including interlaboratory ring trials.

Application of gold immunochromatographic assay strip combined with digital evaluation for early detection of Toxoplasma gondii infection in multiple species.

BACKGROUND: Timely diagnosis of Toxoplasma gondii infection is necessary to prevent and control toxoplasmosis transmission. The gold immunochromatographic assay (GICA) is a means of rapidly detecting pathogen in samples. GICA-based diagnostic methods have been developed to accurately detect pathogens with high sensitivity and specificity, and their application in T. gondii diagnosis is expected to yield good results. METHODS: Colloidal gold test strips were produced using T. gondii C-terminal truncated apical membrane antigen 1 (AMA1C). Colloidal gold-AMA1C and colloidal gold-murine protein conjugate were synthesized under optimal conditions. A nitrocellulose membrane was treated with AMA1C and goat anti-mouse antibody as the test line and control line, respectively. In total, 90 cat serum samples were tested using AMA1C-GICA and a commercial enzyme linked immunosorbent assay (ELISA) kit. The GICA results were digitally displayed using a portable colloidal gold immunochromatographic test strip analyzer (HMREADER). The sensitivity, specificity, and stability of AMA1C-GICA were assessed, and this was then used to examine clinical samples, including 203 human sera, 266 cat sera, and 81 dog sera. RESULTS: AMA1C-GICA had a detection threshold of 1:32 for T. gondii-positive serum. The GICA strips specifically detected T. gondii antibodies and exhibited no reactivity with Plasmodium vivax, Paragonimus kellicotti, Schistosoma japonicum, Clonorchis sinensis, and Schistosoma mansoni. Consequently, 15 (16.7%) positive samples were detected using the AMA1C-GICA and commercial ELISA kits for each of the assays. The receiver-operating characteristic curve showed that GICA had a relative sensitivity of 85.3% and specificity of 92%, with an area under the curve of 98%. After analyzing clinical samples using HMREADER, 1.2%-23.4% of these samples were found to be positive for T. gondii. CONCLUSIONS: This study presents a novel assay that enables timely and efficient detection of serum antibodies against T. gondii, thereby allowing for its early clinical diagnosis. Furthermore, the integration of digital detection using HMREADER can enhance the implementation of GICA.

Comparative performance of ISAGA IgM and ELISA assays for the diagnosis of maternal and congenital Toxoplasma infections: which technique could replace ISAGA IgM?

The ISAGA immunocapture test for the detection of anti-Toxoplasma immunoglobulin M is a manual technique known for its excellent sensitivity and specificity. The purpose of this retrospective, multicenter study was to compare the performances and agreement between ISAGA and other IgM detection techniques before cessation of ISAGA production. The analytic performance of the different tests was evaluated using 1,341 serum samples from adults with positive IgM and negative IgG to Toxoplasma gondii, and 1,206 sera from neonates born to mothers with seroconversion. The agreement between the tests was evaluated on 13,506 adult and 5,795 child serum samples. The sensitivity of Toxo-ISAGA IgM® (adults 98.7%, neonates 63.1%) was similar to that of Platelia Toxo IgM® (adults 94.4%, neonates 64.6%), and significantly higher than Liaison Toxo IgM® (adults 90.6%), Architect/Alinity Toxo IgM® (adults 95.7%, neonates 48.6%), and Vidas Toxo IgM® (adults 81.8%, neonates 17.5%). However, the specificities varied between 24.4% (Platelia Toxo IgM®) and 95.2% (Liaison Toxo IgM®) in adults and were >95% for all tests in neonates. An analysis of the kappa coefficients showed better agreement between ISAGA IgM® and the other tests in children (0.75-0.83%) than in adults (0.11-0.53%). We conclude that, in the absence of Toxo-ISAGA IgM®, the association of a very sensitive technique (Platelia Toxo IgM® or Architect/Alinity Toxo IgM®) and a very specific technique (Vidas Toxo IgM® or Liaison Toxo IgM®) is recommended for IgM detection in adult sera. For neonates, Platelia Toxo IgM® appeared to be the best alternative to replace Toxo-ISAGA IgM®.

Title: Performances comparatives des tests ISAGA IgM et ELISA pour le diagnostic des infections maternelles et congénitales à Toxoplasma : quelle technique pourrait remplacer ISAGA IgM ? Abstract: Le test d'immunocapture ISAGA pour la détection des immunoglobulines M anti-Toxoplasma est une technique manuelle connue pour son excellente sensibilité et spécificité. Le but de cette étude rétrospective et multicentrique était de comparer les performances et la concordance entre l'ISAGA et d'autres techniques de détection d'IgM avant l'arrêt de la commercialisation de l'ISAGA. Les performances analytiques des différents tests ont été évaluées à partir de 1 341 échantillons de sérum d'adultes présentant des IgM positives et des IgG négatives à Toxoplasma gondii, et de 1 206 sérums de nouveau-nés nés de mères présentant une séroconversion. La concordance entre les tests a été évaluée sur 13 506 échantillons de sérum d'adultes et 5 795 sérums d'enfants. La sensibilité de Toxo-ISAGA IgM® (adultes 98,7 %, nouveau-nés 63,1 %) était similaire à celle de Platelia Toxo IgM® (adultes 94,4 %, nouveau-nés 64,6 %) et significativement supérieure à celle de Liaison Toxo IgM® (adultes 90,6 %), Architect/Alinity Toxo IgM® (adultes 95,7 %, nouveau-nés 48,6 %) et Vidas Toxo IgM® (adultes 81,8 %, nouveau-nés 17,5 %). Cependant, les spécificités variaient entre 24,4 % (Platelia Toxo IgM®) et 95,2 % (Liaison Toxo IgM®) chez les adultes et étaient >95 % pour tous les tests chez les nouveau-nés. L'analyse des coefficients kappa a montré une meilleure concordance entre ISAGA IgM® et les autres tests chez les enfants (0,75­0,83%) que chez les adultes (0,11­0,53%). Nous concluons qu'en l'absence de Toxo-ISAGA IgM®, l'association d'une technique très sensible (Platelia Toxo IgM® ou Architect/Alinity Toxo IgM®) et d'une technique très spécifique (Vidas Toxo IgM® ou Liaison Toxo IgM®) est recommandée pour la détection des IgM dans les sérums adultes. Pour les nouveau-nés, Platelia Toxo IgM® apparaît comme la meilleure alternative en remplacement de Toxo-ISAGA IgM®.

Carbon nanospheres dual spectral-overlapped fluorescence quenching lateral flow immunoassay for rapid diagnosis of toxoplasmosis in humans.

Toxoplasmosis is a common zoonotic disease caused by a protozoan parasite Toxoplasma gondii (Tox), approximately infecting one-third of human populations worldwide. This study developed the carbon nanospheres (CNPs) based dual spectral-overlapped fluorescence quenching lateral flow immunoassay (CNPs-FQLFIA) for detection of Tox antibodies (ToxAbs). The CNPs have been effectively coupled with Tox antigen (ToxAg), which can completely overlap the excitation and emission spectra of europium nanospheres (EuNPs) and CdSe/ZnS quantum dots (QDs) in testing strips of CNPs-QDs-FQLFIA or CNPs-EuNPs-FQLFIA. The sensitivity of CNPs-EuNPs-FQLFIA or CNPs-QDs-FQLFIA was 4 or 8 IU/mL under natural light readout, or both 4 IU/mL ToxAbs under ultraviolet (UV) light readout by the naked eyes, respectively. The limit of detection (LOD) of two types of CNPs-FQLFIA was both 1 IU/mL ToxAbs under UV light by a dry fluorescence analyzer, but no cross-reaction was found with other antibodies. The intra-assay coefficient variation (CV) of both CNPs-EuNPs-FQLFIA and CNPs-QDs-FQLFIA was less than 8%, while the inter-assay CV was less than 14%, respectively. The correlation coefficient (R2) of CNPs-EuNPs-FQLFIA or CNPs-QDs-FQLFIA to measure the different concentrations of ToxAbs spiked serum samples was 0.99712 and 0.99896, respectively. The CNPs-FQLFIA presented a characteristics of 94.3% sensitivity, 100% specificity and 98% accuracy for detection of ToxAbs in clinical serum samples. In conclusion, CNPs-FQLFIA with EuNPs or QDs fluorescence reporter was an easy, rapid, sensitive, precise and quantitative assay for detecting Tox antibodies in human blood samples.

Genetic diversity and seroprevalence of Toxoplasma gondii in COVID­19 patients; a first case-control study in Iran.

BACKGROUND: Toxoplasmosis is a serious or life-threatening disease in immunosuppressed patients and pregnant women. This study examined the likely association between Toxoplasma gondii infection and COVID-19 patients with moderate illness. METHODS: Seventy blood samples were collected from patients at the Health Reference Laboratory of Tabriz, Northwest Iran from April 2021 to September 2021. In addition, 70 healthy subjects of the same age (37 ± 15 years) and sex distribution were ethnically matched. Sera samples were examined for the detection of anti-Toxoplasma antibodies using ELISA. Nested-PCR targets were amplified based on the B1 and GRA6 genes. GRA6 amplicons were subjected to sequencing and phylogenetic analysis. RESULTS: The seroprevalence of toxoplasmosis based on IgG titer was 35.7% in the COVID­19 patients and 27.1% in the control group, representing not to be associated with the Toxoplasma seropositivity in COVID­19 patients (P = 0.18) compared to healthy subjects. Anti-T. gondii IgM was not found in any of the patients and healthy individuals. According to PCR amplification of the B1 and GRA6 genes, the frequency of T. gondii in COVID-19 patients was 14.2% (10/70). However, no T. gondii infection was detected in the healthy group. The CD4+T cell count was relatively lower in toxoplasmosis-infected patients (430-450 cells/mm3) than in control group (500-1500 cells/mm3). High genetic diversity (Hd: 0.710) of the type I strain of T. gondii was characterized in the patients. Present results showed that consumption of raw vegetables and close contact with stray cats can increase the transmission of T. gondii to COVID-19 patients (P < 0.01). CONCLUSIONS: The current study revealed that T. gondii type I infection is unequivocally circulating among the COVID-19 patients in Tabriz; However, no significant association was observed between the occurrence of Toxoplasma and the severity of COVID-19. To make more accurate health decisions, multicenter investigations with a larger sample size of different ethnic groups of the Iranian population are needed.

Pre-Transplant Seroprevalence, Associated Factors, and Post-Transplant Incidence of Toxoplasma gondii Infection Among Heart Transplant Recipients in Japan.

BACKGROUND: Among solid organ transplant (SOT) recipients, heart transplant (HT) recipients are at a higher risk of Toxoplasma gondii infection. As Toxoplasma seroprevalence varies by geographic location, updated local epidemiology is essential to guide preventive and therapeutic strategies. However, the Toxoplasma seroprevalence and incidence of post-transplant toxoplasmosis among SOT recipients in Japan are unknown. METHODS: We performed a single-center retrospective observational study at an HT center in Tokyo, Japan. All HT recipients aged ≥18 years between 2006 and April 2019 were included. We reviewed patient charts and conducted a questionnaire survey to investigate the risk factors for infection. RESULTS: Among 105 recipients included in the study, 11 (10.5%) were seropositive before transplant. Ninety-five recipients (90.5%), including all pre-transplant seropositive recipients, answered the questionnaire. The recipients who had lived in Okinawa (odds ratio [OR] 7.5 [95% CI 1.42-39.61]; P = .032) and who reported raw-meat eating habits (OR 4.64 [95% CI 1.04-23.3]; P = .021) were more likely to be seropositive. None of the patients developed symptoms of toxoplasmosis. The post-transplant incidence of other major adverse outcomes was not significantly different according to the pre-transplant serostatus. CONCLUSIONS: About 10% of HT recipients at an HT center in Tokyo were seropositive for Toxoplasma pre-transplant, and none developed symptomatic toxoplasmosis post-transplant on trimethoprim-sulfamethoxazole. The history of raw meat consumption was associated with seropositivity; therefore, avoiding it might be recommended for HT recipient candidates.

Metagenomic Next-generation Sequencing May be a Tool for Timely Diagnosis of Seronegative and Primary Toxoplasma Infection After Allogeneic Hematopoietic Stem Cell Transplantation: A Case Report and Literature Review.

We report a case of Toxoplasma gondii ( T. gondii ) antibody seronegativity in a 14-year-old boy with a primary infection of T. gondii after allogeneic hematopoietic stem cell transplantation for acute T-cell lymphoblastic leukemia who was rapidly diagnosed through metagenomic next-generation sequencing of peripheral blood as well as clinical manifestations. He was successfully cured with timely administration of trimethoprim-sulfamethoxazole due to early diagnosis.

Could PLATELIA Toxo IgM be the new gold standard for the serological diagnosis of congenital toxoplasmosis: a French multicenter study.

To assess the performance of PLATELIA Toxo IgM (Bio-Rad) and Toxo ISAGA (BioMérieux) to detect anti-Toxoplasma IgM in infants at risk of congenital toxoplasmosis, a retrospective multicenter study was conducted comparing serological results obtained in the framework of routine diagnosis work-up for congenital toxoplasmosis. All infants born to mothers infected with T. gondii during pregnancy from 2010 to 2020 with at least 6 months of serological follow-up were included (n = 1,010). One thousand ten cases were included, of which 250 infants (24.75%) had congenital toxoplasmosis. A total of 1039 sera were included. The concordance between the two techniques was 96%, with kappa coefficient of 0.87, showing an almost perfect agreement between ISAGA and PLATELIA. Cumulative sensitivity and specificity were 73.2% and 99.5.% and 74.8% and 100% for ISAGA and PLATELIA, respectively. The mean time to detect IgM using ISAGA and PLATELIA tests was 6.9 ± 20.1 days and 5.6 ± 14.7 days, respectively not significant (ns). Finally, the sensitivity of ISAGA and PLATELIA to detect IgM antibodies in infected neonates at 5 days of life was 62% and 64%, respectively. Performances of PLATELIA Toxo IgM assay were comparable to the gold standard ISAGA. This enzyme-linked immunosorbent assay is suitable for routine serology for the diagnosis of congenital toxoplasmosis in newborns. IMPORTANCE This study will help clinical microbiologists to chose an alternative serological method for the neonatal diagnosis of congenital toxoplasmosis, once the gold standard technique ISAGA will be withdrawn next year.

A Diagnostic Challenge: Toxoplasmosis Masquerading as Tonsillar Neoplasm.

This case report elucidates an uncommon manifestation of toxoplasmosis characterized by an ulcerative oropharyngeal lesion and cervical lymphadenopathy, which intriguingly simulated a tonsillar neoplasm. The patient, a 28-year-old immunocompetent woman, reported symptoms such as a persistent sore throat, unilateral neck pain, and otalgia. Despite the initial clinical impressions, a diagnostic left subtotal tonsillectomy revealed no malignancy but marked acute and chronic inflammation. A comprehensive investigation subsequently indicated a recent primary infection with Toxoplasma gondii, as evidenced by the presence of high IgM antibodies and low IgG avidity. This unique case underlines the significance of incorporating toxoplasmosis into the differential diagnosis of oropharyngeal lesions, thereby necessitating a meticulous approach to laboratory testing. Laryngoscope, 134:1741-1743, 2024.

Comparison of B1 and RE 529 gene targets by real time PCR and LAMP assay for diagnosis of toxoplasmosis in pregnant females.

PURPOSE: The aim of this study is to accurately diagnose the presence of toxoplasmosis in pregnant women. In this study we evaluated two gene targets B1 and RE-529 using two different molecular methods i.e., real-time PCR and LAMP. PROCEDURE: A total of 150 blood samples were collected from pregnant women attending the PGIMER outpatient clinic. The serum and Buffy layer were extracted and various serological (ELISA) and molecular tests (qPCR and LAMP) targeting B1 and RE-529 were carried out. FINDING: Out of 150 patients, 32 were seropositive. Amongst which for the RE-529 gene, 18 were LAMP positive and 16 were qPCR positive, while for the B1 gene, 14 were LAMP positive and 13 were qPCR positive. CONCLUSIONS: Molecular methods were more sensitive than serological tests to diagnose congenital toxoplasmosis in antenatal females. Few seronegative patients were reported positive using molecular methods. In addition, LAMP targeting the RE-529 gene is more sensitive than qPCR, and LAMP targets the B1 gene.

Guidelines for the management of Toxoplasma gondii infection and disease in patients with haematological malignancies and after haematopoietic stem-cell transplantation: guidelines from the 9th European Conference on Infections in Leukaemia, 2022.

Patients with haematological malignancies might develop life-threatening toxoplasmosis, especially after allogeneic haematopoietic stem-cell transplantation (HSCT). Reactivation of latent cysts is the primary mechanism of toxoplasmosis following HSCT; hence, patients at high risk are those who were seropositive before transplantation. The lack of trimethoprim-sulfamethoxazole prophylaxis and various immune status parameters of the patient are other associated risk factors. The mortality of toxoplasma disease-eg, with organ involvement-can be particularly high in this setting. We have developed guidelines for managing toxoplasmosis in haematology patients, through a literature review and consultation with experts. In allogeneic HSCT recipients seropositive for Toxoplasma gondii before transplant, because T gondii infection mostly precedes toxoplasma disease, we propose weekly blood screening by use of quantitative PCR (qPCR) to identify infection early as a pre-emptive strategy. As trimethoprim-sulfamethoxazole prophylaxis might fail, prophylaxis and qPCR screening should be combined. However, PCR in blood can be negative even in toxoplasma disease. The duration of prophylaxis should be a least 6 months and extended during treatment-induced immunosuppression or severe CD4 lymphopenia. If a positive qPCR test occurs, treatment with trimethoprim-sulfamethoxazole, pyrimethamine-sulfadiazine, or pyrimethamine-clindamycin should be started, and a new sample taken. If the second qPCR test is negative, clinical judgement is recommended to either continue or stop therapy and restart prophylaxis. Therapy must be continued until a minimum of two negative PCRs for infection, or for at least 6 weeks for disease. The pre-emptive approach is not indicated in seronegative HSCT recipients, after autologous transplantation, or in non-transplant haematology patients, but PCR should be performed with a high level of clinical suspicion.

The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques.

Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

Past and present seroprevalence and disease burden estimates of Toxoplasma gondii infections in Germany: An appreciation of the role of serodiagnostics.

Toxoplasmosis is one of the major foodborne parasitic diseases in Germany, with 49% of its population chronically infected with its causative agent, Toxoplasma gondii. Although the acute disease is usually benign in immunocompetent individuals, it is a threat for immunocompromised patients as well as for fetuses of seronegative mothers. As a result of infection, congenital and ocular toxoplasmosis can have serious lifelong consequences. Here I will highlight the epidemiologic situation, from its past in the two separate parts of Germany, to its unification 30 years ago and up to the present day. The main identified risk factor for infection in Germany is thought to be the consumption of undercooked or raw meat or sausages. However, the relative impact of this risky eating habit as well as that of other risk factors are changing and are discussed and compared to the situation in the Netherlands. Finally, the importance of robust and efficient high-throughput serological assays for obtaining reliable epidemiological data, on which public health decisions can be made, is highlighted. The potential of bead-based multiplex assays, which allow the incorporation of multiple antigens with different analytical properties and thus yield additional information, are described in this context. It illustrates the interdependence of new analytic assay developments and sound epidemiology, a foundation that decades-old data from Germany did not have.

Tissue cysts and serological detection toxoplasmosis among wild rats from Surabaya, East Java, Indonesia.

Background: The protozoan Toxoplasma gondii is the source of zoonosis toxoplasmosis and causes public health problems throughout the world. Environmental contamination by oocysts excreted by cats as definitive hosts affects the spread of this disease. Wild rats as rodents can be used as an indicator of environmental contamination by oocysts, considering that rats have a habit of living in dirty environments and can be infected by oocysts from the environment. Aim: This study aims to detect toxoplasmosis from tissue cysts and serological tests in wild rats as an indicator of environmental contamination in Surabaya. Methods: A total of 100 wild rats collected from Surabaya were collected in five areas (West, East, Central, North, and South of Surabaya) obtained from three trapping locations: housing, dense settlements, and markets. All samples were examined microscopically for parasitological tests through the brain tissue samples, and the serum was examined using the toxoplasma modified agglutination test to detect the presence of IgG and Immunoglobulin M (IgM). Results: This research used 100 wild rat samples, 77 Rattus tanezumi and 33 Rattus norvegicus, with evidence of 31% in serology and active infection with 19% tissue cyst. The results showed that the seroprevalence of T. gondii in wild rats was 31% (30% for IgG and 1% for IgM). Tissue cysts in the rat brain samples tested were 19% (19/100). The IgG prevalence rate in female rats was 25% (8/32), while for males, it was 32.3% (22/68). The highest seropositive IgG from densely populated settlements was 50%, markets were 25.8%, and housing was 12.1%. The highest seropositive IgM from densely populated settlements was 2.8%. Population density and the presence of cats are factors supporting the high seropositive rate at the trapping location. Conclusion: This study revealed that there has been toxoplasmosis contamination in Surabaya with evidence of 31% in serology and active infection with 19% tissue cyst. It is necessary for controlling with surveillance in cats to prevent transmission in humans.

TORCH Infections.

Perinatal and neonatal infections are a significant cause of morbidity and mortality. As such, early recognition and workup when there is clinical concern is essential to supporting affected neonates. This article aims to focus specifically on the effects of toxoplasmosis, rubella, cytomegalovirus, herpes, and other agents (TORCH) infections, discussing epidemiology, diagnostics, and treatment if available. [Pediatr Ann. 2023;52(11):e400-e406.].

Seroepidemiology of toxoplasmosis in pregnant women and detection of infection acquired during pregnancy in Cotonou, Benin.

Assessing the prevalence of toxoplasmosis in pregnant women and the associated risk factors is the first step in defining policy for the prevention of congenital toxoplasmosis in a given population. An epidemiological study was conducted during prenatal consultations at the CHU-MEL of Cotonou (Benin) between September 2018 and April 2021 and recruited 549 pregnant women to determine the seroprevalence and potential factors associated with Toxoplasma gondii infection. Toxoplasma gondii IgG/IgM antibodies were detected using an enzyme-linked fluorescence assay (ELFA) technique, an IgG avidity test and an IgG/IgM comparative Western blot to diagnose the maternal toxoplasmosis serological status, the possibility of an infection acquired during pregnancy and congenital infection, respectively. Concomitantly, the participants answered a questionnaire investigating potential risk factors. Toxoplasmosis seroprevalence was estimated at 44.4% (95% CI 40.3-48.6) and the factors significantly associated with T. gondii seropositivity were: age over 30 years, multigravid women and contact with cats. The possibility of an infection acquired during the periconceptional period or the first trimester of pregnancy concerned six women [1.1% (95% CI 0.5-2.0)]. However, due to the low rate of serological controls in seronegative women, a significant proportion of women first tested during the 3rd trimester of pregnancy, and an insufficient sample size, the incidence of primary infection during pregnancy could not be determined. No cases of congenital transmission occurred in the newborns from the suspected cases of primary infection.

Title: Séroépidémiologie de la toxoplasmose chez la femme enceinte et détection de l'infection contractée pendant la grossesse à Cotonou, Bénin. Abstract: L'évaluation de la prévalence de la toxoplasmose chez la femme enceinte et des facteurs de risque associés est la première étape pour définir une politique de prévention de la toxoplasmose congénitale dans une population donnée. Une étude épidémiologique a été menée lors des consultations prénatales au CHU-MEL de Cotonou (Bénin) entre septembre 2018 et avril 2021 et a recruté 549 femmes enceintes pour déterminer la séroprévalence et les facteurs potentiels associés à l'infection à Toxoplasma gondii. Les anticorps IgG / IgM de T. gondii ont été détectés à l'aide d'une technique ELFA, du test d'avidité IgG et du Western blot comparatif IgG / IgM pour diagnostiquer respectivement le statut sérologique de la toxoplasmose maternelle, la possibilité d'une infection acquise pendant la grossesse et l'infection congénitale. Parallèlement, les participants ont répondu à un questionnaire portant sur les facteurs de risque potentiels. La séroprévalence de la toxoplasmose a été estimée à 44,4 % (IC 95 % 40,3­48,6) et les facteurs significativement associés à la séropositivité pour T. gondii étaient l'âge supérieur à 30 ans, la multigravidité et les contacts avec les chats. La possibilité d'une infection acquise pendant la période périconceptionnelle ou le premier trimestre de la grossesse concernait six femmes [1,1 % (IC 95 % 0,5­2,0)]. Cependant, en raison du faible taux de contrôles sérologiques chez les femmes séronégatives, d'une proportion importante de femmes testées pour la première fois au cours du 3ème trimestre de la grossesse et d'une taille d'échantillon insuffisante, l'incidence de la primo-infection pendant la grossesse n'a pas pu être déterminée. Aucun des enfants nés des six femmes suspectes de primo-infection en cours de grossesse n'a présenté d'infection congénitale.

Is Childhood Immune Thrombocytopenia Associated With Acquired Toxoplasmosis? An Unusual Case of Infection That Led to Acute ITP in a Greek Male Toddler and Implications for Guidelines.

INTRODUCTION: Childhood immune thrombocytopenic purpura (ITP) is a heterogeneous immune-mediated process triggered by infections, vaccines, allergies and parasites. Currently, there is little evidence in the literature beyond case reports of an association with Toxoplasma gondii (T. gondii). METHODS: The authors describe the unusual case of an earlier healthy 2.5-year-old Greek boy who developed acute ITP with a life-threatening platelet count a few days after a T. gondii infection. Evidence for the infection onset was found incidentally 3 months after the initial admission to the hospital and only after any other plausible cause of thrombocytopenia was excluded, according to diagnosis guidelines. RESULTS: The boy underwent 3 intravenous immunoglobulin treatments within a trimester, a period during which his alarming platelets count levels led to housebound activities. A quite slow recovery was only ignited after the third treatment, which was administered in conjunction with a mild antibiotic medication for the T. gondii infection. Full recovery was obtained 9 months after the initial admission, although the boy's potential scored high in clinical prediction models for developing transient ITP. CONCLUSION: There is a need for more research on ITPs with no obvious cause to investigate a causal association with toxoplasmosis. Currently, testing for diseases of greater rarity and of higher diagnostic cost than T. gondii is included in the ITP guidelines. Hence, routinely testing for toxoplasmosis when considering potential childhood ITP triggers and infection treatment complementary to treating the ITP might be the key to accelerating the healing process and improving the quality of life of otherwise confined children.