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See detailPlace of PCR methods in malaria diagnosis
Klein, Ségolène; Hayette, Marie-Pierre ULg; Melin, Pierrette ULg et al

Conference (2004, October)

Background: Gold-standard method for malaria diagnosis is microscopic examination of Giemsa stained thick and thin blood smears. This method is cheap and simple but fastidious and requires experienced ... [more ▼]

Background: Gold-standard method for malaria diagnosis is microscopic examination of Giemsa stained thick and thin blood smears. This method is cheap and simple but fastidious and requires experienced microscopists. In recent years, molecular biology techniques have been applied with success in the microbiology field because of their great sensitivity and specificity. The aim of this study is the evaluation of Polymerase Chain Reaction (PCR) in the detection of low parasitaemia and mixed infections. Methods: A total of 191 blood samples were included in the study. They were collected from patients admitted to hospital because of suspicion of malaria infection, and distributed as follows: 105 from Liege (Belgium), 42 from Lubumbashi (Democratic Republic of Congo), and 44 from Cayenne (French Guiana). Two PCR techniques targeting the small sub-unit rRNA gene of Plasmodium were tested in comparison with microscopy. The real-time PCR was specific of Plasmodium sp. and the semi-nested multiplex PCR was able to detect each of the four species. Results: The real-time PCR sensitivity was 97% and 100% for multiplex PCR. The specificity of both techniques was 96%. Multiplex PCR detected 2 mixed infections that were missed by microscopy. In 4 cases, both PCR techniques permit to detect parasitaemia after treatment while microscopy was already negative. In one case, parasite DNA was detected by PCR one day before the microscopy became positive. Conclusions: Both PCR techniques presented the same detection limit. The PCR methods had a better sensitivity than microscopy. They detected P. falciparum and P. vivax respectively 7 and 6 days after beginning of treatment. Multiplex PCR allowed species identification and mixed infection determination that could confirm and complete the microscopic examination. Real-time PCR was quicker than nested PCR and could be used for screening in addition to the gold-standard method [less ▲]

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See detailIn vitro drug sensitivity and clinical aspects of Plasmodium falciparum malaria in African children
Carme, Bernard; Gay, Frederic; Ndounga, Mathieu et al

in Tropical Medicine and Parasitology : Official Organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft für Technische Zusammenarbeit (1995), 46(4), 270-274

In vitro Plasmodium falciparum drug sensitivity was investigated in 115 brazzavillians children, between 1 year and 10 years of age. On the basis of clinical aspects, four groups were constituted: Group 1 ... [more ▼]

In vitro Plasmodium falciparum drug sensitivity was investigated in 115 brazzavillians children, between 1 year and 10 years of age. On the basis of clinical aspects, four groups were constituted: Group 1: 39 asymptomatic school children, Group 2: 16 children with uncomplicated malaria, Group 3: 40 with severe but not pernicious malaria and Group 4: 20 with pernicious malaria. The drugs tested were chloroquine (CQ), quinine (QN) and mefloquine (MQ). The sensitivity level was assessed by a 48-hour in vitro maturation test involving the uptake of tritiated hypoxanthine, the initial blood level of parasite being > or = 0.1% in all cases. For QN and MQ, the median IC50 values showed no significant difference related to clinical status, age or parasitaemia levels. For CQ, the proportion of resistant strains and the 50 inhibitory concentration (IC50) values were greater in the cases of children hospitalised for malaria but there were no differences related to clinical severity of these hospitalised children nor, within each group, to the age or parasitaemia levels. The percentage of subjects with an IC50 value greater than the 90 percentile of the IC50 of the asymptomatic group, which we propose as the severity index related to chemoresistance, was 15% for uncomplicated malaria, 38% for severe but non-pernicious forms and 35% for pernicious malaria. The IC50 for QN was significantly higher in CQ-resistant strains and there was a positive correlation for CQ vs QN and for QN vs MQ. [less ▲]

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See detailChimiosensibilité in vivo et in vitro de Plasmodium falciparum à Brazzaville (Congo)
Chandenier, Jacques; Ndounga, Mathieu; Carme, Bernard et al

in Santé : Cahiers d'Etude et de Recherches Francophones (1995), 5(1), 25-29

Various projects were launched in 1993 to monitor the chemosensitivity of Plasmodium falciparum in Congo. Resistance of 34 strains in Brazzaville to chloroquine, quinine and mefloquine and of 35 to ... [more ▼]

Various projects were launched in 1993 to monitor the chemosensitivity of Plasmodium falciparum in Congo. Resistance of 34 strains in Brazzaville to chloroquine, quinine and mefloquine and of 35 to halofantrine was investigated in an in vitro survey using an isotopic micro test. The resistance rates were 61.8, 14.7, 3.0 and 0.0% respectively. Thus, the chemoresistance which first appeared in 1990 is confirmed and is stable in the population. This finding was further confirmed by a parallel in vitro analysis of sensitivity to chloroquine in Brazzaville. A chloroquine monitoring network is now being established throughout the country based on simplified WHO tests of 100 asymptomatic schoolchildren conducted every six months. The first results in 1993, from three Southern regions indicate that parasites are found in 20 to 60% of cases seven days after a standard 3 day treatment with 25 mg/kg, according to the region. The results of in vitro and in vivo tests are very variable. Indeed, the value of such results for these tests for national monitoring is questionable: a more reliable system of identifying true therapeutic failures would be better suited. [less ▲]

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See detailFive cases of infection due to Scedosporium apiospermum
Hayette, Marie-Pierre ULg; Peguet, C.; de Bièvre, Claude et al

Poster (1995)

We report five cases of infection due to Scedosporium apiospermum and his teleomorph Pseudallescheria boydii (Ascomycete), diagnosed in the University Amiens Hospital. The most important clinical ... [more ▼]

We report five cases of infection due to Scedosporium apiospermum and his teleomorph Pseudallescheria boydii (Ascomycete), diagnosed in the University Amiens Hospital. The most important clinical, epidemiological and diagnostic aspects a summurized in the next table. Nb year sex age job clinical statute site mycology (direct culture histology) Ampho B (S or R) TT 1 1992 F 31 secretary sinusitis sinus + S. apiospermum + S surg 2 1993 F 8 schoolgirl cystic fibrosis lung + P. boydii NR R physio 3 1993 M 66 pensioner corticoid cut/scut + S. apiospermum + R keto 4 1994 M 54 farmer corticoid sinus + P. boydii + S surg 5 1994 M 69 pensioner corticoid scut + S. apiospermum NR R surg ampho B: amphotericin B; tt: treatment; histo: histology; S or R: sensitive or resistant; surg: surgery; NR: non realized; physio: physiotherapy, cut/scut: cutaneous/subcutaneous. keto: ketoconazole. Epidemiology. The five cases concern french people living in Picardy (France). All have risk factors: corticotherapy, cystic fibrosis, sinusitis. The average age is 46 years with a sex-ratio 1,5. All the people are living in a rural area and one of them is farmer. The mode of contamination is probably the respiratory tract for the cases 1, 2 and 4. For the third one, the patient reported pricks during a walk in forest. The case 5 occured after forearm surgery. Clinical aspects. The young woman had a past of bacterial sinusal infection with posterior discharge since one year. Unsuccessful treatment With antibiotics and corticoids could explain the development of the fungus. The young girl affected by cystic fibrosis had no modification of the respiratory capacity and we concluded to a simple colonisation of the respiratory tract. For the case 5, the infection by Scedosporium didn't explain the death of the patient three days later. No case led to disseminated infection even for the patients under long-term corticotherapy. Mycology. The teleomorph, P. boydii was observed in only two cases with development of cleistothecia after several weeks of culture at room temperature. In all cases, direct examination showed numerous hyphae excepted the case 4, which showed numerous fungal cells with rare hyphae. Three of the five strains were "in vitro" resistant to amphotericine B and all were sensitive to ketoconazole. Treatment. The treatment were successful in all cases. Conclusion. These five cases of infection due to S. apiospermum are very similar to those described in the literature. Surgery seems to be the best treatment possible. All the strains exhibited low pathogenic power that proves the opportunistic character of this fungus. [less ▲]

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See detailEtude épidémiologique de la toxoplasmose chez les femmes enceintes à Amiens (Picardie): nécessité d'une enquête nationale
Carme, Bernard; Lenne, Evelyne; Tirard, Véronique et al

in Journal de Pédiatrie et de Puériculture (1995), 8(1), 761-763

Il s'agit d'une enquête réalisée à Amiens du 1er mars 1993 au 28 février 1994 chez les femmes venant d'accoucher pour évaluer leur niveau d'immunité vis à vis de la toxoplasmose et leur connaissances des ... [more ▼]

Il s'agit d'une enquête réalisée à Amiens du 1er mars 1993 au 28 février 1994 chez les femmes venant d'accoucher pour évaluer leur niveau d'immunité vis à vis de la toxoplasmose et leur connaissances des mesures préventives. L'analyse des données s'est faite grâce au locigiel Epidemio sur un total de 987 femmes. La séroprévalence est de 58% et l'analyse montre que c'est la consommation de viande peu ou pas cuite et la présence d'un chat dans l'entourage qui sont les 2 facteurs de risque majeurs. La quasi totalité des femmes a pu citer au moins 2 moyens de prévention efficaces et les reccommandations sont largement suivies. En conclusion, les résultats sont rassurants. Cependant, une enquête nationale serait nécessaire pour permettre une évaluation sur le plan national. [less ▲]

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See detailAmibiase intestinale sévère d'origine européenne?
Carme, Bernard; Viot, J.; Hayette, Marie-Pierre ULg et al

in Médecine et Maladies Infectieuses (1995), 25(5),

A case of severe amebic colitis in a 21 year-old french patient is described. It raises the problems of modality and place of contamination, Entamoeba histolytica levels of virulence and differenciation ... [more ▼]

A case of severe amebic colitis in a 21 year-old french patient is described. It raises the problems of modality and place of contamination, Entamoeba histolytica levels of virulence and differenciation between E. histolytica and E. dispar. [less ▲]

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See detailThree cases of infection due to Scedosporium apiopermum
Hayette, Marie-Pierre ULg; Peguet, Chantal; de Bièvre, Claude et al

Poster (1994, October)

We report three cases of infection dur to Scedosporium apiospermum in Amiens university hospital. Two cases concerned two immunosuppressed patient under corticoid tretament, but the third case concerned a ... [more ▼]

We report three cases of infection dur to Scedosporium apiospermum in Amiens university hospital. Two cases concerned two immunosuppressed patient under corticoid tretament, but the third case concerned a young healthy woman with chronic sinusitis. [less ▲]

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See detailDisseminated toxoplasmosis in a patient under methotrexate
Hayette, Marie-Pierre ULg; Claisse, Jean-François; Tribout, B. et al

Poster (1994, October)

A 60 years old patient treated by methotrexate and prednisolone for rheumatoid arthritis, was admitted at the hospital for pulmonary infection refractory to antibiotics. After a few days he was ... [more ▼]

A 60 years old patient treated by methotrexate and prednisolone for rheumatoid arthritis, was admitted at the hospital for pulmonary infection refractory to antibiotics. After a few days he was transferred to the intensive care unit for pancytopenia. Direct examination of bone marow revealed the presence of toxoplasma into polymorphonuclears and monocytes. However, in spite of a specific treatment the patient died two days later of disseminated toxoplasmosis. [less ▲]

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See detailDrug sensitivity and clinical aspects of Plasmodium falciparum malaria in african children
Carme, Bernard; Gay, Frédéric; Tirard, Véronique et al

Conference (1994, October)

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See detailEtude comparative de deux modes de révélation des cultures de toxoplasme en culture cellulaire
Hayette, Marie-Pierre ULg; Elies, Rosen; Chetaille, Eric et al

Poster (1994, June)

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See detailFaisabilité de la PCR dans la détection du développement de Toxoplasma gondii en culture cellulaire MRC5.
Chetaille, Eric; Hayette, Marie-Pierre ULg; Puy, H. et al

Poster (1994, June)

OBJECTIF Etudier la faisabilité de la technique PCR pour la détection du développement de Toxoplasma gondii sur milieu cellulaire à partir des culots et des surnageants de cultures sur fibroblastes ... [more ▼]

OBJECTIF Etudier la faisabilité de la technique PCR pour la détection du développement de Toxoplasma gondii sur milieu cellulaire à partir des culots et des surnageants de cultures sur fibroblastes humains MRCS. METHODES Les surnageants de culture et les cellules correspondant à différents inoculums sont prélevés après un délai variant de 24 à 120h. La technique PCR utilisée comporte une extraction à l'iodure de sodium, l'amplification d'une séquence anonyme TGRI E spécifique de T.gondii (191 paires de bases) et une révélation après électrophorèse sur gel d'agarose-bromure d'éthidium en· UV. La positivité des cultures est controlée parallèlement par immunofluorescence à l'aide d'un anticorps monoclonal antiP30 (Sanofi Pasteur) réalisée sur les culots cellulaires des mêmes cultures. RESULTATS La positivité et la cohérence des résultats erl PCR (positivité de tous les inoculums > plus faible inoculum donnant un résultat positif), aussi bien à partir des culots cellulaires que des surnageants, démontrent la faisabilité de la méthode avec en particulier l'absence d'inhibiteur de la Taq polymérase dans le milieu de culture. CONCLUSION Le développement des toxoplasmes sur milieu cellulaire peut être détecté par la technique PCR. La version sur surnageant de culture à l'intérêt de laisser intact les cellules et de témoigner du développement réel des toxoplasmes. [less ▲]

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See detailEtude épidémiologique de la toxoplasmose chez les femmes enceintes à Amiens (Picardie). Nécessité d'une enquête nationale
Carme, Bernard; Lenne, Evelyne; Tirard, Véronique et al

in Médecine et Maladies Infectieuses (1994), 24

L'étude avait pour but d'évaluer le niveau d'immunité vis à vis de la toxoplasmose des femmes venant d'accoucher à Amiens entre mars 1993 et avril 1994 ainsi que leur connnaissance des mesures préventives ... [more ▼]

L'étude avait pour but d'évaluer le niveau d'immunité vis à vis de la toxoplasmose des femmes venant d'accoucher à Amiens entre mars 1993 et avril 1994 ainsi que leur connnaissance des mesures préventives vis à vis de cette parasitose. Cette étude a permis de mettre en évidence la bonne connaissance des mesures préventives vis à vis de la toxoplasmose chez les femmes non immunes. Ceci a permis également de préciser les données épidémiologiques de la toxoplasmose à Amiens. Le choix d'un protocole incluant les femmes au moment de l'accouchement pourrait être complété par celui de femmes en début de grosesse pour éviter d'exclure les interruptions de grossesse. Cette étude devrait déboucher sur une enquête nationale qui permettrait de préciser l'épidémiolgoie de la toxoplasmose en France. [less ▲]

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See detailEvaluation de la chimiorésistance de Plasmodium falciparum. Revue des tests in vitro: méthodologie, intérêts et limites
Hayette, Marie-Pierre ULg; Carme, Bernard

in Publications Médicales Africaines (1994), 129

The chloroquine-resistance of Plasmodium falciparum appeared first in Southeast Asia and South-America during the sixties. It reached further the african continent in 1977, through Kenya and Tanzania. In ... [more ▼]

The chloroquine-resistance of Plasmodium falciparum appeared first in Southeast Asia and South-America during the sixties. It reached further the african continent in 1977, through Kenya and Tanzania. In Africa, the front of the resistance spread from East to West, and now completely covers Black Africa with different degrees of resistance. Rapidly, "in vivo" and "in vitro" tests were developped to assess the sensitivity of P. falciparum strains against the different antimalarial drugs. This paper is a review of existing "in vitro" and "in vivo" tests with their interest and limits. [less ▲]

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See detailHalofantrine resistance in African countries
Carme, Bernard; Gay, Frédéric; Hayette, Marie-Pierre ULg et al

in Lancet (1993), 341

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See detailHistoplasmose africiaine à Histoplasma duboisii (Histoplasma capsulatum var. duboisii): quatorze cas congolais observés en 10 ans (1981-1990)
Carme, Bernard; Hayette, Marie-Pierre ULg; Itoua Ngaporo, A. et al

in Journal de Mycologie Médicale (1993), 3

Fourteen cases of histoplasmosis due to Histoplasma duboisii seen in Congo from 1981 through 1990, are reported: the average age of the patients was 25 years (the youngest being 1,5 y and the oldest 50y ... [more ▼]

Fourteen cases of histoplasmosis due to Histoplasma duboisii seen in Congo from 1981 through 1990, are reported: the average age of the patients was 25 years (the youngest being 1,5 y and the oldest 50y), the M/F ratio was 2,5; most of the patients (10/14) live in a rural area. there was a rather constant frequency of one to three cases per annum from 1981 through 1990. The fisrt congolese AIDS-associated systemic histoplasmosis duboisii case was observed in 1990. Lymph node involvment and mucocutaneous lesions were the most frequent clinical manifestations. In all cases, the disgnosis was established by histological examination. When performed, direct microscopy was always positive (11/11). Isolation of H. duboisii in culture was successful in 7 out of 9 attempts. Two cases of systemic infection were rapidly fatal but a good clinical response was obtained in the other cases wih amphotericin B, which due to intolerance, was replaced by ketoconazole in 2 patients. [less ▲]

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