RESUMEN
No data concerning antiretroviral drug's (ARV) primary resistance mutation rates in Chad are available. We retrospectively analysed frozen-stored dried blood spot samples that were collected from 48 Chadian human immunodeficiency virus (HIV)-1 seropositive patients naïve of ARV. HIV-1 protease and reverse transcriptase genes were successfully sequenced for 24 (60.0%) of the 40 patients displaying a viral load > 1000 copies/ml. Seven (29.2%) displayed mutations conferring resistance against one or more classes of ARV. We evidenced high levels of primary ARV resistance mutations in Chad, but lower than those observed in patients with failure to first-line ARV.
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Fármacos Anti-VIH/uso terapéutico , Farmacorresistencia Viral , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , VIH-1/efectos de los fármacos , Adolescente , Adulto , Fármacos Anti-VIH/farmacología , Chad/epidemiología , Femenino , Infecciones por VIH/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Carga Viral , Adulto JovenRESUMEN
As a therapy or a support to other therapies, despite being largely beneficial to patients in general, transfusion it is not devoid of some risks. In a moderate number of cases, patients may manifest adverse reactions, otherwise referred to as transfusion-associated hazards (TAHs). The latest French 2016 haemovigilance report indicates that 93% of TAHs are minor (grade 1), 5.5% are moderate (grade 2) and 1.6% are severe (grade 3), with only five deaths (grade 4) being attributed to transfusion with relative certainty (imputability of level [or grade] 1 to 3). Health-care providers need to be well aware of the benefits and potential risks (to best evaluate and discuss the benefit-risk ratio), how to prevent TAHs, the overall costs and the availability of alternative therapeutic options. In high-income countries, most blood establishments (BEs) and hospital blood banks (HBBs) have developed tools for reporting and analysing at least severe transfusion reactions. With nearly two decades of haemovigilance, transfusion reaction databases should be quite informative, though there are four main caveats that prevent it from being fully efficient: (ai) reporting is mainly declarative and is thus barely exhaustive even in countries where it is mandatory by law; (aii) it is often difficult to differentiate between the different complications related to transfusion, diseases, comorbidities and other types of therapies in patients suffering from debilitating conditions; (aiii) there is a lack of consistency in the definitions used to describe and report some transfusion reactions, their severity and their likelihood of being related to transfusion; and (aiv) it is difficult to assess the imputability of a particular BC given to a patient who has previously received many BCs over a relatively short period of time. When compiling all available information published so far, it appears that TAHs can be analysed using different approaches: (bi) their pathophysiological nature; (bii) their severity; (biii) the onset scheme; (biv) a quality assessment (preventable or non-preventable); (bv) their impact on ongoing therapy. Moreover, TAHs can be reported either in a non-integrative or in an integrative way; in the latter case, presentation may also differ when issued by a blood establishment or a treating ward. At some point, a recapitulative document would be useful to gain a better understanding of TAHs in order to decrease their occurrence and severity and allow decision makers to determine action plans: this is what this review attempts to make. This review attempts to merge the different aspects, with a focus on the hospital side, i.e., how the most frequent TAHs can be avoided or mitigated.
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Seguridad de la Sangre , Transfusión Sanguínea/normas , Reacción a la Transfusión , Humanos , RiesgoRESUMEN
BACKGROUND: Skin-dedicated ex vivo fluorescence confocal microscopy (FCM) has so far been used to identify cutaneous tumours on freshly excised samples using acridine orange as fluorochrome. AIM: To use FCM for a new indication, namely, the identification of the herpes simplex virus (HSV) in skin lesions, using fluorescent antibodies. METHODS: Six roof samples from skin vesicles suspicious for HSV lesions were incubated with anti-HSV-1 and anti-HSV-2 antibodies coupled with fluorescein isothiocyanate, and examined under skin-dedicated ex vivo FCM. The positive controls were swabs taken from the floor of each vesicle and observed under conventional direct fluorescence assay (DFA) and by viral cultures. Roof samples from three bullae of bullous pemphigoid were the negative controls. RESULTS: Using ex vivo FCM, the samples from the lesions clinically suspicious for HSV infection were seen to be fluorescent after incubation with anti-HSV-1, and were negative after incubation with anti-HSV-2 antibodies. Conventional DFA with an optical microscope and cultures confirmed the presence of HSV-1 infection. CONCLUSIONS: By using fluorescent antibodies to identify precise structures, ex vivo FCM can be used for indications other than tumour identification. More specifically, it can be an additional diagnostic tool for HSV infection.
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Técnica del Anticuerpo Fluorescente , Herpes Simple/diagnóstico , Herpesvirus Humano 1/inmunología , Microscopía Confocal/métodos , Adolescente , Adulto , Anciano , Anticuerpos Antivirales/inmunología , Femenino , Fluoresceína-5-Isotiocianato , Herpes Simple/inmunología , Herpesvirus Humano 1/aislamiento & purificación , Humanos , MasculinoRESUMEN
Typing of human enterovirus (EV) remains a major goal for diagnostic and epidemiological purposes. Whereas sequencing of the VP1 coding region is the reference standard for EV typing, a method relying on sequencing of the VP2 coding region has been proposed as an alternative; however, this has been validated only on cell culture supernatants. To avoid the selection of cultivable strains and to quicken the identification step, a new semi-nested PCR method targeting the VP2 region was developed by use of the CODEHOP strategy. After validation of the method on reference and clinical strains, a total of 352 clinical specimens found to be positive for EV RNA (138 with the GeneXpert EV kit and 214 with the Enterovirus R-gene kit) during a 3-year period (2010-2012) were analysed prospectively for VP2 genotyping. Overall, 204 (58%) specimens were typeable. A higher proportion of throat swab/stool specimens than of cerebrospinal fluid (CSF) specimens was found to be typeable (94 of 142 (66.2%) vs. 83 of 169 (49.1%), respectively, p <0.01 by the chi-square test). Moreover, the median Ct value obtained was lower for typeable specimens than for untypeable specimens (32.20 vs. 33.01, p <0.05, and 25.96 vs. 31.74, p <0.001, for the GeneXpert and R-gene tests, respectively, by the Mann-Whitney-Wilcoxon test). These results suggest that, in cases of EV meningitis, a peripheral specimen (i.e. throat swab or stool) that is susceptible to exhibiting a higher viral load should be used in preference to CSF for identifying the causative EV genotype by use of the VP2 typing method without cell culture isolation.
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Proteínas de la Cápside/genética , Infecciones por Enterovirus/virología , Enterovirus/clasificación , Enterovirus/genética , Técnicas de Genotipaje/métodos , Reacción en Cadena de la Polimerasa/métodos , Líquido Cefalorraquídeo/virología , Genotipo , Humanos , Datos de Secuencia Molecular , Faringe/virología , Estudios Prospectivos , ARN Viral/genética , Sensibilidad y Especificidad , Análisis de Secuencia de ADNRESUMEN
The novel SPF10 real-time PCR assay allows the simultaneous amplification and detection of the HPV target. That way, LiPA analysis of the HPV-negative samples can be avoided, reducing workload and cost. This study aims to evaluate the performance of the SPF10 real-time PCR in combination with the LiPA assay for HPV detection and typing in cervical samples. Thirty-nine cervical samples were subjected to the SPF10 conventional PCR in combination with the LiPA assay. Subsequently, the SPF10 real-time PCR was performed to enable the comparison between the SPF10 conventional and the real-time PCR results. In case of discrepancy, the samples were subjected to the CLART HPV2 assay. As a result, 27 out of 39 samples were identified as HPV-positive by the SPF10 real-time PCR and were genotyped further by the LiPA assay. Twenty samples (74.1%) showed an absolute agreement between the conventional and real-time SPF10 PCR (concordant), three (11.1%) displayed additional or fewer types (compatible), two (7.4%) did not show any similarity between both assays (discordant) and the remaining two (7.4%) were LiPA-negative. The two assays showed an excellent strength of agreement for individual (κ=0.932) and multiple genotype detection (κ=0.834). In conclusion, the two SPF10 PCR methods are comparable. Therefore, the SPF10 real-time PCR with subsequent LiPA could be used for the detection and genotyping of HPV in cervical samples.
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Cuello del Útero/virología , Técnicas de Diagnóstico Molecular/métodos , Papillomaviridae/clasificación , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/diagnóstico , Infecciones por Papillomavirus/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Femenino , Humanos , Papillomaviridae/genéticaRESUMEN
BACKGROUND: The risk of hepatitis C virus (HCV) transmission during assisted reproductive techniques (ARTs) is still disputed and no report concerning its prospective evaluation is available. METHODS: The aim of this 4-year follow-up multicentre study that enrolled 86 HCV-serodiscordant couples was to determine whether a sperm-processing method was able to reduce levels of HCV in semen and the risk of HCV transmission to the newborn. All the men were chronically infected by HCV and 10 of them by human immunodeficiency virus. A total of 181 seminal plasmas and 153 sperm fractions were tested for the presence of HCV RNA. RESULTS: HCV RNA tested positive in 20.4% of the seminal samples. All of the 153 final sperm fractions tested negative for HCV. The detection of HCV RNA in semen was significantly correlated with a high viral load in blood (P < 0.05). The presence of HCV RNA in seminal plasma impaired neither semen parameters nor ART issue. From the 58 couples enrolled effectively in an ART programme, 24 pregnancies and 28 newborns were obtained. All of them tested negative for HCV RNA in blood. CONCLUSION: These results emphasize the safety of the semen-processing method. The negligible risk of transmitting HCV reduces the value of the systematic analysis of HCV RNA in seminal fractions prior to ART. Since use of this analytical procedure involves the freezing of semen, its avoidance would result in an increase in sperm quality and reduce the need to perform intracytoplasmic sperm injection techniques.
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Hepacivirus/aislamiento & purificación , Hepatitis C/transmisión , Hepatitis C/virología , Semen/virología , Espermatozoides/virología , Adulto , Femenino , Hepacivirus/metabolismo , Humanos , Inseminación Artificial , Masculino , Persona de Mediana Edad , Estudios Prospectivos , ARN Viral/análisis , ARN Viral/sangre , Técnicas Reproductivas Asistidas , Donantes de TejidosRESUMEN
In aged-care facilities, gastroenteritis outbreaks are responsible for big trouble in the management of cares to the elderly. In November 2002, a gastroenteritis outbreak was observed in 5 of the 6 wards of the geriatric hospital La Charité, University Hospital of Saint-Etienne, France, with an attack rate of 38.5% in the elderly (70 infected from 182 patients) and of 26.0% in the nursing staff (40 infected from 154 agents). The outbreak lasted 30 days with a peak corresponding to 79.8% of the cases between the 11(th) and the 20(th) of November. The first cases were observed in the two short-term-care wards; then, the outbreak spread rapidly to 3 of the 4 long-term care units. Health care workers were contaminated later than the elderly (P < 0.001 by Kruskal-Wallis test). A self-administered questionnaire was documented by most of the nursing staff; the most frequently observed clinical symptoms in this population were nausea (82.5%), abdominal pain (80.0%), diarrhoea (70.5%), asthenia (67.5%) and vomiting (62.5%). Thirty-five percent of the health care workers ceased their work. The causative agent of the gastroenteritis was identified by RT-PCR in the stools of 5 aged persons as a norovirus close to the Lordsdale strain (genogroup II). These findings illustrate the respective role of elderly and health care workers in the spread of the gastroenteritis outbreak inside the geriatric hospital.
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Infecciones por Caliciviridae/epidemiología , Brotes de Enfermedades , Gastroenteritis/epidemiología , Gastroenteritis/virología , Norovirus , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Geriatría , Hospitales Especializados , Humanos , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: Heterosexual human immunodeficiency virus (HIV) transmission implies the crossing of the vaginal mucosa by virions present in the semen, potentially using Langerhans cells as transporters. The recruitment of these cells in the mucosa is mediated by the chemokine macrophage inflammatory protein 3alpha (CCL20). The aim of this study was to evaluate the capacity of the semen to induce Langerhans cell recruitment via the production of CCL20 by vaginal epithelial cells. METHODS AND RESULTS: Using a vaginal epithelium model based on the SiHa cell line and human seminal plasma, we demonstrated that semen enhanced the production of CCL20. This secretion was regulated by the nuclear factor-kappaB intracellular signalling pathway. Fractionation of the seminal plasma indicated that the secretion of CCL20 was stimulated by high molecular weight compounds present in semen. Migration assays demonstrated that secreted CCL20 was able to promote the recruitment of Langerhans cell precursors (LCps), which remain permissive to X4 and R5 HIV infection. CONCLUSIONS: Our results demonstrate that epithelial cells respond to factors present in semen by secreting CCL20, leading to the enhancement of LCp recruitment. These data argue in favour of the implication of epithelial cells in the heterosexual transmission of HIV.
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Quimiocinas CC/metabolismo , Infecciones por VIH/transmisión , Células de Langerhans/inmunología , Proteínas Inflamatorias de Macrófagos/metabolismo , Semen/inmunología , Enfermedades Virales de Transmisión Sexual/transmisión , Vagina/inmunología , Movimiento Celular/inmunología , Quimiocina CCL20 , Células Epiteliales/inmunología , Femenino , Infecciones por VIH/inmunología , Humanos , Masculino , FN-kappa B/metabolismo , Enfermedades Virales de Transmisión Sexual/inmunología , Transducción de Señal , Vagina/citologíaRESUMEN
This cross-sectional study aimed to investigate, during a short period between 2000 and 2001, in a large population of patients with chronic hepatitis C, the epidemiological characteristics of hepatitis C virus (HCV) genotypes in France. Data from 26 referral centres, corresponding to 1769 patients with chronic hepatitis C were collected consecutively during a 6-month period. HCV genotyping in the 5'-non-coding region (NCR) was performed in each center using the line probe assay (LiPA, in 63% of cases), sequencing (25%) or primer-specific polymerase chain reaction (PCR) (12%). HCV genotypes 1a, 1b, 2, 3, 4, 5, non-subtyped 1 and mixed infection were found in 18, 27, 9, 21, 9, 3, 11 and 1% of our population, respectively. HCV genotype distribution was associated with gender, age, source and duration of infection, alanine aminotransferase (ALT) levels, cirrhosis, alcohol consumption, hepatitis B virus (HBV) and human immunodeficiency virus (HIV) coinfection. In multivariate analysis, only the source of infection was the independent factor significantly associated with genotype (P = 0.0001). In conclusion, this study shows a changing pattern of HCV genotypes in France, with i.v. drug abuse as the major risk factor, an increase of genotype 4, and to a lesser extent 1a and 5, and a decrease of genotypes 1b and 2. The modification of the HCV genotype pattern in France in the next 10 years may require new therapeutic strategies, and further survey studies.
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Hepacivirus/clasificación , Hepacivirus/genética , Adulto , Estudios de Cohortes , Femenino , Francia/epidemiología , Genotipo , Hepacivirus/aislamiento & purificación , Hepatitis C/epidemiología , Hepatitis C/fisiopatología , Hepatitis C/virología , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Reacción en Cadena de la Polimerasa , ARN Viral/genéticaRESUMEN
OBJECTIVE: The authors had for aim to study the distribution of HIV-1 subtypes in a cohort of HIV-1 positive patients in the University hospital of Saint-Etienne, France, and to describe the epidemiological characteristics of patients infected with a non-B subtype strain. DESIGN: An epidemiological study was made on 271 HIV-1 positive patients followed up in the Infectious Diseases Department over 20 years. All patients sample were subtyped by serotyping and some samples were also tested by genotyping. RESULTS: Two hundred and sixty-four patients (191 men and 73 women) were found infected by an HIV-1 strain belonging to the M group. After combining serotyping and genotyping results, 195 patients were found infected by a B subtype and 69 by a non-B subtype. Most of the latter strains belonged to an A subtype or related ones. The following factors were shown to be linked to an infection by a non-B strain: being born abroad, having contracted the infection though heterosexual practice, and being a woman. The incidence of non-B strains increased regularly over time (to reach more than 40% in 2003). This progression was especially noted for men born in France with risky sexual behaviour. CONCLUSION: These results indicate that more than 40% of HIV-1 new cases detected in the Saint-Etienne area are related to non-B strains and that strains of A and related subtypes are common in the local population with risky sexual behaviour.
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Infecciones por VIH/virología , VIH-1/clasificación , Femenino , Francia , Genotipo , Humanos , Masculino , Prevalencia , SerotipificaciónRESUMEN
Prevalence of hepatitis G virus among Tunisian blood donors Hepatitis G virus (GBV-C/HGV) is a recently identified virus which occurs worldwide. The prevalence of GBV-C/HGV in Tunisia has not been previously studied. We aimed to assess the prevalence of GBV-C/HGV infection in Tunisian blood donors. A total of 912 blood donors were tested for anti-E2 antibodies of GBV-C/HGV by enzyme-linked immunosorbent assay and 600 were tested by reverse transcriptase polymerase chain reaction. GBV-C/HGV RNA was found in 5.3% of the sample and HGV antibodies occurred in 4.9%. A correlation was noticed between GBV-C/HGV infection and hepatitis C virus (P = 0.006). The prevalence of GBV-C/HGV is similar to that reported worldwide.
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Donantes de Sangre/estadística & datos numéricos , Virus GB-C , Hepatitis Viral Humana/epidemiología , Hepatitis Viral Humana/virología , Adolescente , Adulto , Distribución por Edad , Comorbilidad , ADN Viral/análisis , ADN Viral/genética , Ensayo de Inmunoadsorción Enzimática , Femenino , Virus GB-C/genética , Virus GB-C/inmunología , Infecciones por VIH/complicaciones , Infecciones por VIH/epidemiología , Anticuerpos Antihepatitis/sangre , Hepatitis B/complicaciones , Hepatitis B/epidemiología , Hepatitis C/complicaciones , Hepatitis C/epidemiología , Hepatitis Viral Humana/sangre , Hepatitis Viral Humana/complicaciones , Hepatitis Viral Humana/diagnóstico , Humanos , Masculino , Tamizaje Masivo/métodos , Persona de Mediana Edad , Vigilancia de la Población , ARN Viral/análisis , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Riesgo , Estudios Seroepidemiológicos , Túnez/epidemiologíaRESUMEN
Prevalence of hepatitis G virus among Tunisian blood donors Hepatitis G virus [GBV-C/HGV] is a recently identified virus which occurs worldwide. The prevalence of GBV-C/HGV in Tunisia has not been previously studied. We aimed to assess the prevalence of GBV-C/HGV infection in Tunisian blood donors. A total of 912 blood donors were tested for anti-E2 antibodies of GBV-C/HGV by enzyme-linked immunosorbent assay and 600 were tested by reverse transcriptase polymerase chain reaction. GBV-C/HGV RNA was found in 5.3% of the sample and HGV antibodies occurred in 4.9%. A correlation was noticed between GBV-C/HGV infection and hepatitis C virus [P = 0.006]. The prevalence of GBV-C/HGV is similar to that reported worldwide
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Adolescente , Adulto , Distribución por Edad , Comorbilidad , ADN Viral , Resumen en Inglés , Donantes de SangreRESUMEN
Hemodialysed patients are recognised as a group at increased risk of infection with hepatitis C virus (HCV). The aim of this study was to determine the prevalence and incidence of HCV infection among dialysis patients of the east-centre part of Tunisia. Two hundred and seventy-six patients dialysed until 2001 were recruited within seven hemodialysis units located in the cities of Sousse, Monastir and Mahdia. The serum markers of HCV infection were tested over the period of March 2000-December 2002, by a 3rd generation ELISA test for antibodies and by qualitative RT-PCR technique for viral RNA. The prevalence of anti-HCV antibodies and of HCV RNA was 32.6% (90 patients) and 25.7% (71 patients), respectively. Between 1998 and 2002, 20 new infections were documented in five of the seven dialysis units corresponding to an incidence of 2.34% per year, with an average time of contamination after the beginning of dialysis of 4.6 years. If all the infections are assessed to have occurred during dialysis, the density of incidence of HCV contamination was 4.4% per year of dialysis. A high correlation was noticed between the presence of HCV markers in serum and the duration of dialysis (F = 34.15, P < 0.0001). In the absence of other risk factors (transfusion, drug-addiction), these results plead for the nosocomial transmission of the observed HCV infections. A phylogenetic analysis of the E2 hypervariable region of the viral genome is in progress to confirm this assumption.
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Hepatitis C/epidemiología , Diálisis Renal/efectos adversos , Femenino , Hepacivirus/genética , Hepacivirus/aislamiento & purificación , Humanos , Incidencia , Masculino , Prevalencia , ARN Viral/genética , ARN Viral/aislamiento & purificación , Estudios Retrospectivos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Túnez/epidemiologíaRESUMEN
BACKGROUND: In France, assisted reproductive technologies involving a hepatitis C virus (HCV)-infected man requires the cryopreservation of potentially infected semen (in order to establish the presence of HCV), hence the need for a safe and secure storage system. We evaluated the safety of high-security straws for the conservation of semen containing HCV RNA under routine conditions. METHODS: Ionomeric resin (IR) straws were filled with seminal plasma spiked with different concentrations of HCV RNA and sealed using a thermo-solder. After a 4% sodium hypochlorite treatment and/or cryopreservation for 7 days in liquid nitrogen, the outside ends of each straw were rinsed with RNAse-free water. RESULTS: No HCV RNA could be detected in any of the water samples. Additional samples included the rinsing water from straws sealed by thermo-solder and from the heating wire used to cut the end of straws containing HCV-positive semen. The latter samples were found positive for both HCV RNA and the protamine-2 gene expressed by spermatozoa. CONCLUSIONS: These results demonstrate the safety of IR straws, the filling system and the thermo-solder for cryopreservation of semen containing HCV in liquid nitrogen. Decontamination of the straw after sealing and the use of disposable scissors to open the straws are strongly recommended.
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Criopreservación , Hepacivirus/aislamiento & purificación , Técnicas Reproductivas Asistidas , Preservación de Semen , Semen/virología , Criopreservación/instrumentación , Hepacivirus/genética , Humanos , Masculino , ARN Viral/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Irrigación Terapéutica , Microbiología del AguaRESUMEN
Hepatitis C virus (HCV) strains isolated from 68 haemodialysis Tunisian patients exhibiting chronic infection were genotyped targeting the NS5b region of the HCV genome using a prototype assay developed by Bayer HealthCare-Diagnostics (TRUGENE NS5b HCV). The overall results were compared to those obtained with another assay of the same company based on sequencing of the 5' non-coding region (TRUGENE HCV 5'NC genotyping kit). All strains could be typed by the 5'NC typing kit, but only 62 (91; 2%) by the NS5b prototype assay. All the 62 strains typed by both methods exhibited the same pattern at the type level: 57 were type 1, 3 were type 2, and 2 were type 4. At the subtype level, eight strains that gave undetermined results by the 5'NC kit were successfully typed by the NS5b kit; eight additional strains exhibited discrepant results. The overall agreement between the two assays was 74.2% at the subtype level. In conclusion, the NS5b region appears to be much more accurate than the 5'NC region to subtype HCV strains, especially in those isolated from patients attending haemodialysis centres where the subtype distribution suggests frequent nosocomial transmissions.
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Regiones no Traducidas 5'/análisis , Hepacivirus/clasificación , Juego de Reactivos para Diagnóstico , Proteínas no Estructurales Virales/análisis , Regiones no Traducidas 5'/química , Estudios de Evaluación como Asunto , Genotipo , Hepacivirus/genética , Humanos , Túnez , Proteínas no Estructurales Virales/químicaRESUMEN
Many mouse models of human enterovirus disease have been pro- posed, concerning both acute and persistent infection. However, rather paradoxically since the usual way of contamination is fecal-oral, most of them used a systemic route of infection. The aim of the present work was to follow the development of an experimental enterovirus infection and to study the viral persistence at the organ level. Twenty-eight female 3-week old BALB/c mice were infected with 5 x 10(4) TCID(50) of coxsackievirus B3 (CV-B3), Nancy strain, by oral route using a rigid cannula introduced into the stomach. The kinetics of infection was studied by sacrificing 2 animals at different times post infection (from 1 hour to 90 days). The presence of the virus in various organs (small intestine, heart, pancreas, lung, spleen, kidney, liver) was studied by cell culture and RT-PCR. As soon as one hour post infection, the virus was detected in the small intestine. In the heart, the virus was present at 24 and 48 hours post infection by RT-PCR and culture, respectively. At 5 days post infection, all the organs but the liver were found infected. The virus was detected up to 15 days in kidney, 21 days in pancreas, 30 days in lung and spleen, and 45 days in intestine, by both culture and PCR. The heart was still found infected 90 days post infection by both techniques. These results show the dramatic cardiotropism of CV-B3 inoculated by oral route, with a detection of the virus very soon in the course of infection (24 hours) and a persistence of the virus for more than 3 months. The intestine, the initial target of enterovirus infection, can also be considered as a site of viral persistence.
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Infecciones por Coxsackievirus , Modelos Animales de Enfermedad , Animales , Infecciones por Coxsackievirus/transmisión , Infecciones por Coxsackievirus/virología , Femenino , Ratones , Ratones Endogámicos BALB C , BocaRESUMEN
Maternofetal transmission of Toxoplasma gondii was assessed in pregnant guinea-pigs, with a gestational period of 65 +/- 5 days. A total of 56 female guinea pigs was infected by the intraperitoneal route (RH strain), by the oral or the intraperitoneal route (Prugniaud strain; PRU) or by the oral route (76K strain). Inoculation was performed 90 +/- 18 days or 30 +/- 9 days before the onset of gestation or 20 +/- 6 days or 40 +/- 6 days after. Gestational age was determined by a progesterone assay. Parasite loads (fetal brain and liver) were assessed by nested PCR and real-time PCR quantification on Light Cycler was performed with a SYBR Green I technique. The 76K strain appeared to be the most virulent in the model: the neonatal survival rate was 31%, in contrast to 53% and 68% for the PRU and RH strains, respectively. The percentage of survival of neonates for all strains taken together was lower after inoculation at 40 days' gestation (25%) than at 20 days' gestation (77%). Whatever the strain, maternofetal transmission determination was greater with nested PCR (54% for RH, 84% for PRU and 86% for 76K strains) than with real-time quantitative PCR (31% for RH, 66% for PRU and 76% for 76K strains). However, real-time quantitative PCR showed that neonatal parasite load was greater with the cystogenic strains (76K, PRU) and that high hepatic load (> 10000 parasites/g) was often associated with disease severity (11 of 12 cases). Therefore, this technique may provide an important element in understanding this congenital disease.
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Toxoplasmosis Animal/congénito , Toxoplasmosis Animal/transmisión , Animales , Modelos Animales de Enfermedad , Femenino , Cobayas , Transmisión Vertical de Enfermedad Infecciosa , Masculino , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/estadística & datos numéricos , Embarazo , Complicaciones Parasitarias del Embarazo/parasitología , Sensibilidad y Especificidad , Toxoplasma/genética , Toxoplasma/aislamiento & purificación , Toxoplasmosis Animal/complicaciones , Toxoplasmosis Animal/parasitología , Toxoplasmosis Cerebral/complicaciones , Toxoplasmosis Cerebral/parasitología , Toxoplasmosis Cerebral/transmisiónRESUMEN
In France, assisted reproductive technology (ART) for hepatitis C virus (HCV)-infected patients is now subject to strict control after the publication of recent guidelines. Infertile serodiscordant couples (HCV-viraemic men and their seronegative female partners) require special care to carried out in designated 'viral risk' laboratories. Twelve sequential semen samples taken from an HCV chronically infected patient were analysed within 22 months. HCV RNA was detected in all the seminal plasma sampled before antiviral treatment with relatively high viral loads, and in two of the corresponding fractions of motile sperm obtained after a gradient selection, suggesting that a contamination risk by HCV through ART cannot be excluded. When the selection of sperm on a discontinuous gradient was followed by an additional swim-up step, HCV RNA was never detected in the motile sperm suspension that was frozen in highly secure straws. IVF was performed using cryopreserved sperm that tested negative for HCV RNA, resulting in a pregnancy. One month after embryo transfer, testing for HCV RNA and antibodies in the woman gave negative results.
Asunto(s)
Fertilización In Vitro , Hepacivirus/genética , Hepatitis C/transmisión , ARN Viral/análisis , Espermatozoides/virología , Adulto , Antivirales/administración & dosificación , Femenino , Hepatitis C/tratamiento farmacológico , Hepatitis C/prevención & control , Humanos , Infertilidad Femenina/terapia , Masculino , Embarazo , Resultado del Embarazo , Semen/virología , Carga ViralRESUMEN
OBJECTIVE: To investigate the presence of the genome of GB virus C/hepatitis G virus (GBV-C/HGV) in semen and saliva from HIV-1-infected men. METHODS: Samples of blood from 33 men seropositive for HIV-1 were tested for the presence of GBV-C/HGV markers of infection, RNA by RT-PCR, and anti-E2 antibodies by ELISA, respectively. The cell-free fractions of seminal fluid and saliva samples of the patients with positive blood samples for GBV-C/HGV RNA or anti-E2 antibodies were then analyzed for the presence of the RNA of this virus. In addition, six semen samples and 11 saliva samples from GBV-C/HGV-negative men were tested. RESULTS: The GBV-C/HGV RNA tested by RT-PCR was recovered from blood in 11 patients of 33 (33.3%), and the antibodies to E2 envelope protein were detected in six patients (18.2%). Since no patient was positive for both markers, the overall prevalence of GBV-C/HGV infection was 51.5% in the studied population. Four-all belonging to the homosexual risk group-of the 17 men with markers to GBV-C/HGV in blood were found to be positive for GBV-C/HGV RNA in mucosal samples: two of them exhibited genomic RNA in both semen and saliva, and two others were positive for semen only. The absence of inhibitors of the PCR technique was confirmed in all mucosal fractions found negative for GBV-C/HGV RNA, except for one saliva sample and one seminal fluid sample. CONCLUSION: These results confirm the high prevalence of GBV-C/HGV infection in patients infected with HIV-1 by sexual exposure and the presence of GBV-C/HGV RNA in seminal fluid and saliva of men with markers of this virus in the blood, suggesting that mucosal fluids could be a potential source for the spread of the GBV-C/HGV infection.
Asunto(s)
Virus GB-C/aislamiento & purificación , Infecciones por VIH/virología , VIH-1 , Saliva/virología , Semen/virología , Antígeno 12E7 , Antígenos CD/análisis , Antígenos CD/inmunología , Moléculas de Adhesión Celular/análisis , Moléculas de Adhesión Celular/inmunología , Virus GB-C/genética , Humanos , Masculino , ARN Viral/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVES: Report of epidemiological, clinical and virological data collected from the prospective surveillance of febrile episodes observed in aged residents of a long-stay care unit of 33 beds, at the University Hospital of Saint-Etienne, during the 1997-1998 winter season. METHODS: Systematic collection of clinical and biological data from febrile patients (> or = 38 degrees C) on a form, including virological findings obtained from a nasal swab and paired serum specimens. RESULTS: From 38 patients (37 of them having been vaccinated against influenza in October 1997), 18 febrile episodes were recorded in 16 subjects, including 3 respiratory syncytial virus infections and a late-occurring outbreak (March 1998) of influenza due to a A/H3N2 strain (15 cases, 14 of them virologically confirmed). No death was noted after the influenza outbreak. In 8 of the 9 tested patients with influenza, "protective" titres of antibodies directed towards the hemagglutinin of the vaccinal strain were present by radial hemolysis test three months before the beginning of the outbreak. During the influenza outbreak, the attack rate of symptomatic infection was 45.5% in elderly and 47.5% in healthcare workers (mainly unvaccinated). The occurrence of the first cases in the latter suggests their possible role in the transmission of the virus to the aged. CONCLUSION: This study underlines the epidemic circulation of multiple respiratory viruses during the same winter season in long-stay care facilities, the occurrence of clinical influenza infections in vaccinated patients exhibiting protective antibody titres and the role of unvaccinated healthcare workers in the propagation of influenza in institutionalised aged.