Hélène Fricker-Hidalgo

Detection of Circulating Aspergillus fumigatus Galactomannan: Value and Limits of the Platelia Test for Diagnosing Invasive Aspergillosis

ABSTRACT The effectiveness of galactomannan detection with the Platelia test was evaluated in a prospective study of 3,327 sera from 807 patients. The specificity was 99.6% (748 of 751 cases). For the groups of patients with proven and probable invasive aspergillosis, the sensitivity was 50.0% (17 of 34 cases). The disappointing sensitivity associated with the presence of rare false-positive cases underlines the limits of this test.

Determination of anti–Toxoplasma gondii immunoglobulin G avidity: adaptation to the vidas system (bioMérieux)

The determination of specific anti-Toxoplasma gondii IgG avidity has been proposed to improve the determination of the date of toxoplasmic seroconversion in pregnant women. In this study, we adapted this serological technique to the Vidas system (bioMérieux) using 6 M urea as the dissociating agent. We studied 356 sera, including 42 sequential sera from sero-conversions in pregnant women. Our results show that the test is easy to use, and that an avidity index higher than 0.300 allows the exclusion of a recent infection acquired less than 4 months before serum sampling.

Diagnosis of toxoplasmosis after allogeneic stem cell transplantation: results of DNA detection and serological techniques.

BACKGROUND The biological diagnosis of toxoplasmosis after allogeneic hematopoietic stem cell transplantation (HSCT) is based on the detection of Toxoplasma gondii DNA in blood specimens or other samples. Serological testing is used mainly to define the immunity status of the patient before HSCT. The aim of our study was to examine the performance of polymerase chain reaction (PCR) and serological techniques in the diagnosis of toxoplasmosis after HSCT. METHODS Seventy patients underwent allogeneic HSCT from September 2004 through September 2006. DNA was detected by PCR, and immunoglobulin G and immunoglobulin M were detected by enzyme-linked immunosorbent assay. RESULTS The results of immunoglobulin G detection before allogeneic HSCT were positive in 40 (57.1%) of the patients and negative in 30 (42.9%). After HSCT, 57 patients (81.4%) had test results that were negative for immunoglobulin M and had negative results of DNA detection, without toxoplasmosis infection. Four patients (5.7%) had at least 4 samples with positive PCR results and/or test results positive for immunoglobulin M against T. gondii; toxoplasmosis was then confirmed by clinical symptoms. Nine patients (12.9%) with positive PCR results and 1 or 2 samples with test results negative for immunoglobulin M were considered to have asymptomatic T. gondii infection. Reactivation of latent infection was the cause of toxoplasmosis in 3 of the 4 patients, and toxoplasmosis occurred as a primary infection in 1 patient. The detection of specific anti-T. gondii immunoglobulin M was the only biological evidence of toxoplasmosis in 2 patients, and samples were positive for immunoglobulin M before PCR was performed in 1 patient. CONCLUSIONS Thus, after HSCT, all patients were at risk for toxoplasmosis; all patients who receive HSCTs should be followed up with biological testing that combines PCR and serological techniques.

The placenta: a main role in congenital toxoplasmosis?

Systemic infections, such as toxoplasmosis, acquired during pregnancy can lead to placental infection and have profound effects on the mother-to-child relationship and the success of pregnancy. Placental permeability to Toxoplasma gondii is a main parameter that determines parasite transmission to the foetus, and the use of antibiotics to decrease placental parasite load and prevent congenital toxoplasmosis has been suggested for decades. Although parasitological examination of the placenta at birth is commonly used to diagnose neonatal congenital toxoplasmosis, this approach can be controversial. Here we argue in favour of placental examination for both diagnostic and epidemiological purposes.

Evaluation of Candida ID, a New Chromogenic Medium for Fungal Isolation and Preliminary Identification of Some Yeast Species

ABSTRACT Candida ID, a new chromogenic medium, allows identification ofCandida albicans (blue colonies) and preliminary identification into a group of four species (pink colonies). In comparison with Albicans ID2 and Sabouraud gentamicin chloramphenicol on 446 fungal strains, Candida ID allowed the isolation of more species than Albicans ID 2 (95.5% versus 91.2%).

Multivariate analysis comparing microbial air content of an air-conditioned building and a naturally ventilated building over one year

Heating, ventilation and air-conditioning (HVAC) may be responsible for the production and spread of airborne microorganisms in office buildings. In order to compare airborne microbiological flora in an air-conditioned building with that in a naturally ventilated building, eight sets of measurements were made over a 1-year period. Concurrently with other environmental measurements, air samples were collected in each building, from three offices and from the outdoor air, using the Andersen single-stage sampler. Three different media were used to culture fungi, staphylococci and mesophilic bacteria. Multivariate analysis revealed a group of offices more contaminated than others, and a marked seasonal variation in fungal concentrations. A comparison of mean levels of microorganisms measured in the two buildings showed that the air microbial content was significantly higher and more variable in the naturally ventilated building than in the air-conditioned building. Moreover, in the naturally ventilated building, the interior fungal content was strongly dependent on the outdoor content, while in the air-conditioned building fungal concentrations remained constant despite significant variations measured outside. This was confirmed by a statistical comparison of the correlation coefficients between indoor and outdoor concentrations. No difference was observed regarding gaseous pollutants and temperature, but relative humidity was significantly higher in the air-conditioned building. The effect of HVAC was to prevent the intake of outdoor particles and to dilute the indoor concentrations. These results are consistent with the presence of high-efficiency filters and a steam humidifier in the HVAC system under study.

Serodiagnosis of recently acquired Toxoplasma gondii infection in pregnant women using enzyme-linked immunosorbent assays with a recombinant dense granule GRA6 protein.

Indirect immunoglobulin G (IgG) and IgM enzyme-linked immunosorbent assays (ELISAs) with a recombinant GRA6 protein of Toxoplasma gondii were developed and evaluated for accurate diagnosis of recently acquired infection in pregnant women. According to the results from Toxoplasma serodiagnostic tests, women were classified into 3 groups representing acute (group I), chronic (group II), or no Toxoplasma infection (group III). To discriminate group I from group II sera, the GRA6-IgG-ELISA reached sensitivity and specificity of 87.5% and 94.1%, respectively. Although 22 (91.7%) of 24 group I sera were positive by the GRA6-IgM-ELISA, only 1 (2.9%) of 34 group II sera scored positive. The GRA6-IgM-ELISA displayed a meaningful correlation with Vidas Toxo IgM and exhibited higher specificity (97.1%) than Euroimmun IgM ELISA (88.2%) (Euroimmun, Lübeck, Germany) for detection of recent infection. These results demonstrate that IgG and IgM ELISA with rGRA6 are useful to identify and discriminate recent from past Toxoplasma infection in pregnant women.

Recombinant Allergens Combined with Biological Markers in the Diagnosis of Allergic Bronchopulmonary Aspergillosis in Cystic Fibrosis Patients

ABSTRACT Allergic bronchopulmonary aspergillosis (ABPA) is a frequent complication in cystic fibrosis patients. The diagnosis remains difficult and requires a combination of clinical, radiological, biological, and mycological criteria. The aim of this study was to analyze the added value of two recombinant antigens, rAspf4 and rAspf6, associated with the detection of specific IgG; precipitins; total IgE; and Aspergillus fumigatus in sputum for the diagnosis of ABPA. In a retrospective study, we determined the specific IgE responses to these recombinants in 133 sera of 65 cystic fibrosis patients. We selected an average of five serum samples from each of the 17 patients with ABPA (13 proven and 4 probable ABPA) and from 3 patients with Aspergillus bronchitis and rhinosinusitis. One serum sample for the 45 patients without ABPA was tested. The sensitivity of specific IgE detection against rAspf4 calculated per patient (92.3%) was significantly higher (P < 0.05) than that of rAspf6 (53.8%). When rAspf4 IgE detection was associated with anti-Aspergillus IgG enzyme-linked immunosorbent assay (ELISA) and precipitin detection, the sensitivity rose to 100%. The specificities of rAspf4 and rAspf6 IgE detection were 93.7% and 91.6%, respectively. Other diagnostic criteria had slightly lower specificities (87.5% for anti-Aspergillus IgG ELISA, 89.6% for precipitins, 84.4% for total IgE, and 85.0% for positive A. fumigatus culture in sputum). In conclusion, this retrospective study showed the relevance of rAspf4 IgE detection, in combination with other biological markers (Aspergillus IgG ELISA, precipitins, and total IgE), for improving the biological diagnosis of ABPA.

PRENATAL DIAGNOSIS OF CONGENITAL TOXOPLASMOSIS: COMPARATIVE VALUE OF FETAL BLOOD AND AMNIOTIC FLUID USING SEROLOGICAL TECHNIQUES AND CULTURES

The prenatal diagnosis of congenital toxoplasmosis is mainly based on biological tests performed on fetal blood and amniotic fluid. We studied the performance of neonatal diagnosis procedures and the results of fetal blood and amniotic fluid analysis. Of 127 women who contracted toxoplasmosis and underwent prenatal diagnosis, the postnatal serological follow‐up was long enough to definitively diagnose congenital toxoplasmosis in 19 cases and to exclude it in 27 cases. Prenatal diagnosis allowed the detection of 94.7 per cent (18/19) of the infected fetuses. The sensitivities of tests in amniotic fluid and fetal blood were equivalent, 88.2 per cent (15/17) and 87.5 per cent (14/16), respectively. In fetal blood, biological techniques were positive in 12/16 cases and in 2/16 cases, serological tests were the only positive sign. The specificities of tests in amniotic fluid and fetal blood were respectively 100 per cent (23/23) and 86.3 per cent (19/22) (three false‐positive serological results). These results, added to the lower morbidity of amniocentesis compared with cordiocentesis, might lead to cordocentesis being abandoned in the prenatal diagnosis of congenital toxoplasmosis.

Toxoplasma Seroconversion with Negative or Transient Immunoglobulin M in Pregnant Women: Myth or Reality? A French Multicenter Retrospective Study

ABSTRACT Classically, Toxoplasma infection is associated with high levels of specific IgM antibody and a rise in specific IgG levels 1 to 3 weeks later. Atypical IgG seroconversion, without IgM detection or with transient IgM levels, has been described during serologic follow-up of seronegative pregnant women and raises difficulties in interpreting the results. To evaluate the frequency and the characteristics of these atypical cases of seroconversion, an investigation was conducted within the French National Reference Center for Toxoplasmosis, from which 26 cases collected from 12 laboratories belonging to the network were identified. The aim of this work was to retrospectively analyze the results of serologic testing, the treatments administered, and the results of prenatal and postnatal follow-up for these women. In each case, IgG antibodies were detected using both screening and confirmatory tests. IgM antibodies were not detected in 15 cases, and the levels were equivocal or low-positive in 11 cases. The IgG avidity results were low in 16 cases and high in one case. Most of the pregnant women (22/26) were treated with spiramycin from the time that IgG antibodies appeared until delivery. Amniotic fluid was analyzed for Toxoplasma gondii DNA by PCR in 11/26 cases, and the results were negative in all cases. Congenital toxoplasmosis was ruled out in 12/26 newborns. There was no abnormality observed at birth for 10 newborns and no information available for 4 newborns. In conclusion, when the interpretation of serological results is so difficult, it seems cautious to initiate treatment by spiramycin and to follow the pregnant women and their newborns.