Toxoplasmosis in Transplant Recipients, Europe, 2010-2014

Transplantation activity is increasing, leading to a growing number of patients at risk for toxoplasmosis. We reviewed toxoplasmosis prevention practices, prevalence, and outcomes for hematopoietic stem cell transplant (HSCT) and solid organ transplant (SOT; heart, kidney, or liver) patients in Europe. We collected electronic data on the transplant population and prevention guidelines/regulations and clinical data on toxoplasmosis cases diagnosed during 2010-2014. Serologic pretransplant screening of allo-hematopoietic stem cell donors was performed in 80% of countries, screening of organ donors in 100%. SOT recipients were systematically screened in 6 countries. Targeted anti-Toxoplasma chemoprophylaxis was heterogeneous. A total of 87 toxoplasmosis cases were recorded (58 allo-HSCTs, 29 SOTs). The 6-month survival rate was lower among Toxoplasma-seropositive recipients and among allo-hematopoietic stem cell and liver recipients. Chemoprophylaxis improved outcomes for SOT recipients. Toxoplasmosis remains associated with high mortality rates among transplant recipients. Guidelines are urgently needed to standardize prophylactic regimens and optimize patient management

The role of DNA microarrays in Toxoplasma gondii research, the causative agent of ocular toxoplasmosis

Ocular toxoplasmosis, which is caused by the protozoan parasite Toxoplasma gondii, is the leading cause of retinochoroiditis. Toxoplasma is an obligate intracellular pathogen that replicates within a parasitophorous vacuole. Infections are initiated by digestion of parasites deposited in cat feces or in undercooked meat. Parasites then disseminate to target tissues that include the retina where they then develop into long-lived asymptomatic tissue cysts. Occasionally, cysts reactivate and growth of newly emerged parasites must be controlled by the host’s immune system or disease will occur. The mechanisms by which Toxoplasma grows within its host cell, encysts, and interacts with the host’s immune system are important questions. Here, we will discuss how the use of DNA microarrays in transcriptional profiling, genotyping, and epigenetic experiments has impacted our understanding of these processes. Finally, we will discuss how these advances relate to ocular toxoplasmosis and how future research on ocular toxoplasmosis can benefit from DNA microarrays

[Real-time PCR in the diagnosis of toxoplasmosis: the way to standardisation?]

Severity of toxoplasmosis is highly correlated to the immune status of the infected individual. Foetus and immunocompromised patient are mostly at risk to develop life threatening forms. In this situation, serological diagnosis gives poor information. DNA detection using polymerase-chain-reaction technology (PCR) has significantly improved the management of this disease. Even so, the growing number of conventional PCR assays has finally led to variable performance results. Real-Time PCR (RT-PCR) in toxoplasmosis has been developed since 2000. This new technology can improve standardisation. Moreover, quantification of parasitic load in samples becomes possible. This review describes the main RT-PCR procedures actually under use and the studies comparing different target genes. The effective benefit of quantification is also discussed. Reducing number of procedures and more systematic external quality control should be considered, in order to improve reliability in PCR results, which has undoubtedly become a major tool in toxoplasmosis diagnosis

Toxoplasma gondii in human astrocytes in vitro: interleukin (IL)-12 and IL-10 do not influence cystogenesis.

Interleukin (IL)-12, IL-10, and interferon (IFN)-gamma are major cytokines involved in the immune response against Toxoplasma gondii. Nevertheless, the role of IL-12 and IL-10 in the control of parasite replication and cytogenesis is not known yet, whereas the importance of IFN-gamma is documented. Furthermore, it is of paramount importance to study the interaction between T. gondii and cells from the central nervous system, e.g., astrocytes. In this study, we report that IL-12 and IL-10 have no effect on penetration, replication, or cystogenesis of the T. gondii Prugniaud strain in human astrocytes in vitro and do not antagonize the role of IFN-gamma on cystogenesis

Evaluation of a new packaging process for non-autoclavable endoscopes: results for the first 100 microbiological samples

International audienc

Disseminated toxoplasmosis with pulmonary involvement after heart transplantation.

We report a case of pulmonary toxoplasmosis after heart transplant despite the prophylactic anti-toxoplasmic treatment that was given but was not sufficient to prevent toxoplasmosis. However, the patient survived thanks to early diagnosis confirmed by polymerase chain reaction on blood and by serological techniques, and early treatment

Adapting a conventional PCR assay for Toxoplasma gondii detection to real-time quantitative PCR including a competitive internal control.

We have developed a quantitative PCR assay (LightCycler* using the pair of primers JW58 and JW59 for the detection of the 35-fold repeated B gene of oxoplasma gondii. This real-time PCR, using fluorescence resonance energy transfert (FRET) hybridization probes, allows the quantification of . gondii with several technical requirements not previously described: i) an internal amplification control (co-amplified in a single tube with the same primers), ii) Uracil-N-Glycosylase and iii) a standard curve corresponding to a serial dilution from a calibrated suspension of T. gondii ranging from 40 to 4.106( )parasites in one ml of amniotic fluid (1 to 105( ) . gondii/PCR). In artificial samples, one parasite could be detected if at least three reactions were performed

Evaluation of a new packaging process for non-autoclavable endoscopes: results for the first 100 microbiological samples

International audienc

Comparative Assessment of a Commercial Kit and Two Laboratory-Developed PCR Assays for Molecular Diagnosis of Congenital Toxoplasmosis

International audienceToxoplasmosis is a worldwide infection that may cause severe disease and is regarded as a serious health problem in France. Detection of the parasite by molecular methods is crucial for diagnosing the disease. The extreme diversity of methods and performances of Toxoplasma PCR assays makes the use of commercial PCR kits an attractive alternative, as they offer a chance for standardization. We compared the performances of three molecular methods for the detection of Toxoplasma gondii DNA in amniotic fluid: a commercial method using nested PCR and two laboratory-developed methods, one using conventional PCR and the other one real-time PCR. This evaluation was based upon a T. gondii DNA serial dilution assay, three amniotic fluid samples spiked with T. gondii at different concentrations, and a clinical cohort of 33 amniotic fluid samples. The T. gondii DNA serial dilution assay showed a much lower sensitivity for the commercial kit than for the laboratory-developed methods. Moreover, out of 12 proven congenital toxoplasmosis cases, 91.7% were detected by the laboratory-developed assays, whereas only 50% were detected by the commercial kit. A lack of sensitivity of the method, partly due to the presence of PCR inhibitors, was the main drawback of the commercial method. This study emphasizes that commercial PCR diagnostic kits do not systematically perform better than carefully optimized laboratory-developed methods. There is a need for thorough evaluation of such kits by proficient groups, as well as for performance standards that commercial kits can be tested against to improve confidence in those selected by health care providers