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P Truc

Publications and source records attributed to P Truc.

At least 19 recordsLinked to original sources

Genetic characterization of Trypanosoma brucei gambiense and clinical evolution of human African trypanosomiasis in Côte d'Ivoire.

Human African trypanosomiasis is a parasitic infection caused by protozoa belonging to Trypanosoma brucei subspecies. The clinical evolution of this disease is complex and might be because of the parasite itself, as genetic diversity has been observed in T. brucei ssp. We investigated the relationship between the genetic diversity of trypanosomes and the diversity of clinical patterns in Côte d'Ivoire. We studied clinical sleeping sickness cases, and genetically analysed the trypanosomes isolated from these patients. An important genetic monomorphism among stocks isolated in Côte d'Ivoire was observed by using various markers: isoenzymes electrophoresis, random amplified polymorphism DNA and PCR of microsatellite sequences. At the same time, the diversity of clinical patterns and evolutions was confirmed by clinical analysis. The existence of an individual susceptibility to disease (human trypanotolerance) should be taken into account even if our genetic conclusions might be distorted because the isolation success rates were particularly poor. In fact, we observed that the isolation success rate varied significantly depending both on the focus of origin (P=0.0002) and on the ethnic group (P=0.0317) of the patient. Further investigations are required in order to study a possible selective impact of the use of the kit for in vitro isolation of trypanosomes as an isolation technique.

Animals↗

IgM quantification in the cerebrospinal fluid of sleeping sickness patients by a latex card agglutination test.

An increased IgM concentration in cerebrospinal fluid (CSF), occurring as a consequence of massive intrathecal IgM synthesis, is a marker of interest for diagnosis of the meningo-encephalitic stage in human African trypanosomiasis. However, in current practice, IgM in CSF is not determined because of the lack of a simple and robust test that is applicable in African rural regions where the disease prevails. We describe the development of a sensitive semiquantitative card agglutination test, LATEX/IgM, for IgM quantification in CSF. The test is simple and fast and the lyophilized reagent remains stable even at 45 degrees C. CSF end-titres obtained with LATEX/IgM parallel the IgM concentrations determined by nephelometry and enzyme-linked immunosorbent assay. Detection of intrathecal IgM synthesis is the most sensitive marker for CNS involvement in sleeping sickness. At a cut-off value of >or= 8, the sensitivity and specificity of LATEX/IgM for intrathecal IgM synthesis are 89.4 and 92.7%. As a consequence, patients with LATEX/IgM end-titres >or= 8 are likely to have intrathecal IgM synthesis, thus central nervous system involvement and therefore should be treated accordingly. Further studies should concentrate on the relationship between the LATEX/IgM end-titres, presence of intrathecal IgM synthesis and occurrence of treatment failures in patients treated with pentamidine.

Animals↗

[Methodology to determine risk zones for sleeping sickness in Côte d'Ivoire by the spatial approach].

Earth observation technology is a rapidly expanding field in which new outlets for sensor data are being found daily. However medical applications are still limited. The main obstacle is the lack of methods to link spatial data to an epidemiological data base within a given disease context. This obstacle was recently overcome in the Sinfra region of Cote d'Ivoire where a number of researchers have been working on human African trypanosomiasis (HAT) for over ten years. A geographical information system was developed to allow testing of various spatial relationships especially between the population density and disease prevalence using various methods of geostatistical analysis. Spot satellite data was then used to draw a map showing population density. The data base was used to validate processing by explaining the presence of errors due mainly to locality. Finally as a first experimental application for this methodology, a map showing the risk of HAT transmission over the entire Sinfra region was drawn based on satellite data.

Cote d'Ivoire↗

[Distribution and spread of human African trypanosomiasis: value of genetic identification of the trypanosomes].

Numerous factors extrinsic to trypanosome populations have been implicated in the distribution and spread of human African trypanosomiasis (HAT), but quantification of these factors has proven difficult. An easier method of monitoring HAT consists of tracking parasites by genetic identification of trypanosomes in hosts and vectors. This method requires distinction between Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense followed by determination of the genotype of each subspecies in the host and vector. The role of vertebrate hosts in the distribution and spread of HAT has been confirmed by genetic identification of trypanosomes. Despite the ever-improving performance of biomolecular techniques, identification must be carried out on natural populations actually circulating in the biological fluids of the host and vector. This precaution is necessary to rule out the biasing effect of in vivo or in vitro isolation of trypanosomes.

Animals↗

Preliminary evaluation of LATEX/T. b. gambiense and alternative versions of CATT/T. b. gambiense for the serodiagnosis of human african trypanosomiasis of a population at risk in Côte d'Ivoire: considerations for mass-screening.

A study was conducted to compare classical card agglutination test for trypanosomiasis (CATT)/T. b. gambiense with CATT-EDTA and LATEX/T. b. gambiense as alternative field tests for serodiagnosis of Human African Trypanosomiasis. The tests were performed on freshly collected blood in an endemic and a low prevalence area in Côte d'Ivoire. Diagnostic performance of each test was assessed using Quantitative Buffy Coat as the parasitological reference and immune trypanolysis as the serological reference test. According to the parasitological data, CATT-EDTA on 10 microl and LATEX/T. b. gambiense on blood diluted 1:4, detecting all confirmed cases with good specificity (respectively 94.6% and 98.1%) yielded better results than the classical CATT did (one false negative and 92.5% specific). However, when immune trypanolysis data and feasibility are taken into account, the classical CATT remains the test of choice for mass screening under the given field conditions.

Agglutination Tests↗

Characterization of Trypanozoon isolates using a repeated coding sequence and microsatellite markers.

Genetic variation of microsatellite loci is a widely used method for linkage analysis, individual identification or inter-population studies. Here we analyse a repeated DNA coding sequence and eleven new microsatellites identified within the Trypanosoma (Trypanozoon) brucei genome. Ninety-seven isolates belonging to the five species and subspecies Trypanosoma evansi, T. equiperdum, T. brucei brucei, T. b. rhodesiense and T. b. gambiense were compared regarding the genetic patterns of these markers. The results reveal a great heterogeneity of the genotypes related to the repeated coding sequence and five microsatellites, some of which show a high degree of polymorphism. This allows us to define group-specific genotypes or alleles; in particular, we show that one specific pattern clearly segregates the human pathogen T. b. gambiense group I.

Amino Acid Sequence↗

Characterization of Trypanozoon isolates using a repeated coding sequence and microsatellite markers.

Genetic variation of microsatellite loci is a widely used method for linkage analysis, individual identification or inter-population studies. Here we analyse a repeated DNA coding sequence and eleven new microsatellites identified within the Trypanosoma (Trypanozoon) brucei genome. Ninety-seven isolates belonging to the five species and subspecies Trypanosoma evansi, T. equiperdum, T. brucei brucei, T. b. rhodesiense and T. b. gambiense were compared regarding the genetic patterns of these markers. The results reveal a great heterogeneity of the genotypes related to the repeated coding sequence and five microsatellites, some of which show a high degree of polymorphism. This allows us to define group-specific genotypes or alleles; in particular, we show that one specific pattern clearly segregates the human pathogen T. b. gambiense group 1.

Journal Article↗

Identification of bloodmeals in haematophagous Diptera by cytochrome B heteroduplex analysis.

We developed a DNA assay for bloodmeal identification in haematophagous insects. Specific host cytochrome B gene sequences were amplified by PCR and classified on the basis of their mobility in a heteroduplex assay. In the blackfly Simulium damnosum s.l. (Diptera: Simuliidae), human cytochrome B DNA sequences were identifiable up to 3 days following ingestion of the bloodmeal. In the tsetse Glossina palpalis (Diptera: Glossinidae) collected from tsetse traps in Ivory Coast, bloodmeals were identified as taken from domestic pigs on the basis of their heteroduplex pattern and DNA sequence. Evidently the cytochrome B sequence shows sufficient interspecific variation to distinguish between mammalian host samples, while exhibiting minimal intraspecific variation. The stability of DNA in bloodmeals, for several days post-ingestion by haematophagous insects, allows PCR-HDA assays to be used reliably for host identification.

Animals↗

Use of polymerase chain reaction in human African trypanosomiasis stage determination and follow-up.

Stage determination of human African trypanosomiasis is based on the detection of parasites and measurements of biological changes in the cerebrospinal fluid (CSF) (concentration of white blood cells > 5 cells per mm3 and increased total protein levels). The patient is treated accordingly. Demonstration of the absence or presence of trypanosomes by the double centrifugation technique is still the only test available to clinicians for assessing treatment success. In this study, however, we evaluate the polymerase chain reaction (PCR) as a tool for assessing the disease stage of trypanosomiasis and for determining whether treatment has been successful. All 15 study patients considered to be in the advanced stage of the disease were PCR positive; however, trypanosomes were demonstrated by double centrifugation in only 11 patients. Of the five remaining patients, who were considered to be in the early stage, PCR and double centrifugation were negative. Following treatment, 13 of the 15 second-stage patients were found to be negative for the disease in at least two samples by PCR and double centrifugation. Two others were still positive by PCR immediately and one month after the treatment. Trypanosome DNA detection using PCR suggested that the two positive patients were not cured but that their possible relapse could not be identified by a search for parasites using the double centrifugation technique. Further evaluation of the PCR method is required, in particular to determine whether PCR assays could be used in studies on patients who fail to respond to melarsoprol, as observed in several foci.

Animals↗

Identification of trypanosomes isolated by KIVI from wild mammals in Côte d'Ivoire: diagnostic, taxonomic and epidemiological considerations.

In Côte d'Ivoire, a comparative study was carried out on 122 wild mammals by parasitological and serological examination and by in vitro isolation of trypanosomes from fresh blood (KIVI). Thirteen isolated stocks were studied by isoenzymes and compared with Trypanosoma congolense and T. brucei bouaflé group reference stocks. Of the 122 animals, only 22 were positive on blood smears while 88 were KIVI positive and 92 were CATT/T. b. gambiense positive. For six stocks identified by isoenzymes as T. congolense, the agreement between ELISA and CATT was good (75%). As compared with CATT, antigen detection ELISA was not satisfactory for T. brucei (20%). Out of 18, 16 stocks represented a separate zymodeme (seven T. congolense and nine T. brucei) and a high genetic heterogeneity was observed. For T. congolense, savanna, kilifi and forest groups were represented by one zymodeme each. The four remaining zymodemes while put into this T. congolense group, were strongly independent of each other. Morphology indicated that those new zymodemes correspond to T. congolense. In the other hand, five new zymodemes fit into T. brucei classification.

Animals↗

A new method for identifying blood meals of human origin in tsetse flies.

A new sensitive technique using the electrophoresis of superoxide dismutase to distinguish between tsetse blood meals of human and non-human origin is described. In Côte d'Ivoire, 602 blood meals were collected; 170 were from man (28.3%), 377 from animals (62.6%) and 55 were unidentified (9.1%) because no pattern was observed. When calculating the index of epidemiological risk, it is strongly correlated with the incidence of sleeping sickness cases.

Animals↗

[Epidemiologic risk of infectious disease transmission in regions of the Ivory Coast].

An epidemiological risk indicator based on purely entomological factors can be used to identify regions which are at higher risk of transmission within the endemic forest zones of Ivory Coast, and can serve to point out biotopes to be treated to make antivector campaigns more effective. In the forests of Ivory Coast, so-called socially open environments which are populated by a great number of ethnic groups who are highly mobile and whose amps are spread over a large area, are particularly vulnerable to epidemics of the disease. Transmission always occurs near water: at rivers, water holes, plantations. By contrast, socially closed societies consisting of a single ethnic group settled in a village are at much lower risk. It seems that human behaviour plays as much a role in transmission dynamics as environmental changes which may be more or less favourable to the vector.

Animals↗

A high prevalence of mixed trypanosome infections in tsetse flies in Sinfra, Côte d'Ivoire, detected by DNA amplification.

The prevalence of various species and subgroups of trypanosomes in the Sinfra area of Côte d'Ivoire was determined using the polymerase chain reaction (PCR). Using this technique to amplify specific satellite DNA targets, it was possible to identify developmental-stage trypanosomes in the midguts and the proboscides of tsetse without expansion of parasite populations. The predominant tsetse species in the area was Glossina palpalis, while G. pallicera and G. nigrofusca were also present. Microscopical examination of 811 non-teneral flies revealed an infection rate of 14% in midguts and/or proboscides. Three subgroups of Trypanosoma congolense (Savannah, Forest & Kilifi), T. simiae, T. godfreyi, West African T. vivax and T. brucei ssp. were identified using PCR. T. congolense Forest was the most abundant of the Nannomonas trypanosomes. Approximately 40% of all infections were mixed, and there was a significantly higher prevalence of apparently mature T. brucei ssp. trypanosomes than has previously been reported. The present study demonstrates that PCR facilitates the easy identification of mature trypanosome infections in tsetse, providing a reliable estimation of trypanosomiasis challenge.

Animals↗

[Serologic diagnosis of human african trypanosomiasis caused by Trypanosoma brucei gambiense. 1--Production and use of rough antigens in ELISA and latex agglutination tests].

The extraction of rough antigens (Ag) from trypanosomes came across a major problem getting of a high number of parasites. Moreover several technics for the extraction of rough antigens are described. Some of them are sophisticated and time consuming. Aiming the obtention of a large number of Ag for this study, inoculation of trypanosomes into Cricetomys gambianus and affinity chromatography of blood allow to collect of large number of trypanosomes. This new technique leads to the extraction of 50 to 60 mg of rough Ag of trypanosomes isolated from infected Cricetomys. In order to set up a specific and sensitive serological diagnosis of gambian human trypanosomiasis, the different rough Ag were used comparatively by ELISA and Latex Agglutination tests. Confirming previous works, the results indicate a high sensitivity and a doubtful specificity for both techniques.

Animals↗