[Emergence of Chagas' disease in Europe: description of the first cases observed in Latin American immigrants in mainland France]. / Emergence de la maladie de Chagas en Europe: à propos des premiers cas observés chez des migrants latino-américains en France métropolitaine.

This article describes the first cases of imported Chagas' disease detected in Paris, France. A total of 18 cases were recorded in two teaching hospitals between 2004 and 2007. There were 12 women and six men with a mean age of 38 years. All patients were Latin American immigrants who had recently arrived in France from Bolivia (Cochabamba and Santa-Cruz departments) 17 cases and from Salvador in 1. Eleven patients presented an asymptomatic indeterminate form of the chronic disease. Seven presented chronic Chagas cardiomyopathy including two with severe symptoms requiring placement of a pacemaker. Obtaining serological tests to confirm the diagnosis was difficult. All except one patient who was older than 50 years were treated with benznidazole. Based on these findings, the main priorities for management imported Chagas' disease in France are improvement of serological diagnosis and prevention of vertical transmission.

[Chagas disease in chronic phase outside the endemic area. The diagnostic tools]. / Maladie de Chagas en phase chronique hors zone d'endémie. Les outils diagnostiques.

The diagnosis of Chagas disease during the chronic phase is based on serology. Outside South America the use of two methods is recommended by WHO. A third method must be available for inconclusive results but there is no gold standard. A pilot study of screening in 254 Bolivian people living in the Paris area (France) was made. Serological study was performed using IIF and three Elisa, Elisa Cruzi (BioMérieux Brésil), BioElisa Chagas (Bio-kit), and Chagatest Elisa recombinante v. 3.0 (Wiener Lab). 165 patients were negative, 69 positive and 20 inconclusive. PCR-based assays appear to have a better sensitivity than parasitological methods, but not more than 70% that do not justify their use for primary testing. There are no standardized and commercial assays. The primer pairs based on the nuclear sequence TCZ1-TCZ2 seems to be the more specific (no cross reaction with others Trypanosomatidae) and the most sensitive with the strains of the two lineage of Trypanosoma cruzi. PCR would have a role in inconclusive serological cases or in the evaluation of treatment failure.

[Experience of targeted screening of Chagas disease in Ile-de-France]. / Expérience de dépistage ciblé de la maladie de Chagas en Ile-de-France.

2009 is marked by the centenary of the discovery by Carlos Chagas of Human American Trypanosomiasis. As a result of international cooperation its incidence has been falling in endemic areas, whereas North America and Europe are witnessing an increase in the number of imported cases. In metropolitan France, 18 such cases were reported between 2004 and 2007. Recently, estimates based on immigration figures have been made and suggest that about 1,500 imported cases can be expected in France. The object of this article is to assess the value of targeted screening of an at-risk population, originally from Latin America and now living in the Ile-de-France (area centred on Paris). The serological techniques employed were indirect immunofluorescence (IIF) and, depending on the case, 2 or 3 Elisa tests (Biomérieux, Biokit and Wiener). Trypanosoma cruzi serology was considered positive when the IIF was superior or equal to 200, or when two Elisa's were > 1, or when the IIF was superior or equal to 100 with at least one Elisa > 1. PCR was performed in 48 cases, which were considered to be positive. The tests were carried out on a voluntary basis after a publicity campaign within the Latin American community in the Ile-de-France. In this article, we present the findings of the first year of screening. Two hundred and fifty-four individuals were screened for Chagas' disease between June 2008 and June 2009. The median age was 33 years [11-63], the male/female ratio 102/152. Overall prevalence of positive serology was 23.6% (60/254). For six patients, the results were classified as "uncertain" (discordant serological tests). Of the seropositive group, 87.4% were Bolivian and 100% presented as a chronic form. Of these, 23.6% presented with functional cardiac manifestations and 22% with gastro-intestinal problems. The PCR was positive in 61% of the seropositive individuals. Clinical evaluation together with other investigations and therapeutic intervention is being carried out at present. These results confirm that metropolitan France is subject to the emergence of Chagas' disease in a non-endemic zone. This confirms the value of screening in at-risk populations, in particular because of the recent broadening of indications for antiparasitic treatment. In addition it is relevant to the prevention of vertical transmission or infection via organ donation, which could arise in France. These results also demonstrate continuing difficulties in the interpretation of serological results and the usefulness of PCR, which might increase sensitivity substantially.

Measures to prevent transfusion-associated protozoal infections in non-endemic countries.

To understand the risk of protozoa transmission by blood is critical as: (i) the world has become globalized with extensive travel, and increased immigration; (ii) blood-borne protozoa is common in inter-Tropical areas; (iii) protozoa develop biological means to escape hosts' immune systems, together with complicated detection, surveillance, and biological testing; and (iv) life threatening-parasites are inadequately controlled by treatment or prevention. This question is relevant in France, with it's non-continental territories, such as French Guiana, located in the Amazon Basin, which is endemic for various Plasmodium ssp. responsible for malaria, and for Trypanosoma cruzi, which is responsible for Chagas disease. In France, specific questioning of blood donors is haphazard despite the increase in population migration over the last three decades: specific questioning must be emphasized and 'at-risk' donors should be identified and subsequently excluded from donation. Donor exclusion alone would only be partially efficient, there is also a need for relevant biological testing of blood donations and in particular for T. cruzi through the CE-marked test to organize a coherent prevention policy: precise studies would thus define which blood donations are subjected to this additional qualifying test when available.

[Post-transfusion parasite transmission: do the present controls fit with the EU directive?]. / Le risque parasitaire transfusionnel : quels contrôles au regard de la directive européenne?

Blood transfusion has become extremely safe regarding the transmission of infectious pathogens, some of them having been responsible for mostly severe complications and a certain loss of confidence from practitioners and patients over the last decades. This may result from the strict observance of ethical principles, of a better medical selection of donors, of technical steps for preparing and qualifying blood components for therapeutic use. The transfusion systems--which vary in their constitution and missions depending on the countries or even regions--have imposed themselves strict security rules and guidelines in industrialized countries. Further, governmental sanitary authorities have set up additional surveillance systems to make the transfusion systems the safest as possible. In addition, the Council of Europe has edicted directives to redefine guidelines to the preparation, use and quality assurance of blood products, that are mandatory in countries of the European Community. Regarding the infectious risks, most recommendations have focused more on the bacterial (immediate) and the viral (mostly delayed) risks than on the parasitic risks because these risks are not only less frequent in industrialized countries, but also far less well known and even much more complex. However, because travel habits and immigrations are increasing fast, most transfusion systems or blood banks must revisit their practices and controls towards hemoparasite transmission by blood transfusion. This review aims at discussing the present controls set up in most industrialized countries and particularly in Europe regarding the risk of post-transfusion transmission of hemoparasites, and the robustness of such controls as well as how these controls may be secured by the European Directive.

[Hepatitis G virus and labile blood products: role of transfusional transmission]. / Virus de l'hépatite G et produits sanguins labiles: rôle de la transmission transfusionnelle.

The GBV-C/HGV (HGV) virus was discovered a few years ago. This virus is known to be parenterally as well as sexually transmitted. However, no study has found some pathogenic roles for HGV so far. In the present study, we aimed to investigate the transmission of HGV by blood components transfused to 284 patients hospitalized in surgery unit in 1995. We tested two parameters of infection in blood components transfused to infected recipients: viral RNA by PCR and anti-E2 antibodies by ELISA. We tried to suspect some potent hepatocyte impacts by assessing the levels of two enzymes in serums: alanine aminotransferase (ALT) and alpha-glutathion S-transferase (alpha-GST). We found that HGV-RNA was detectable in 3.6% of recipients prior to transfusion and 7.5% post-transfusion. For each infected recipient, we retrospectively did a search for HGV-RNA in each transfused blood component, and we found at least one blood component as HGV-RNA-positive for each transfusional infected recipient. Anti-E2 antibody prevalence standing for a former and cured infection was 39.6% in all the recipients. In viremic recipients, ALT levels were mostly normal, while alpha-GST levels were found more commonly elevated than in non-viremic recipients although non-significantly (20% vs. 6.3%; P = 0.07). The present study underlines that HGV transmission is mostly transfusional in surgery units, and that infectiosity of blood components can be anticipated by detection of the viral RNA by PCR. Furthermore, the possible relationship between the serum activity of alpha-GST and the hepatotropism of HGV, although non-admitted as pathogenic, should be investigated.

Multicentric evaluation of the DiaMed enzyme-linked immunosorbent assay malaria antibody test for screening of blood donors for malaria.

BACKGROUND: The risk of malaria transmission by blood transfusion is critical due to extensive travel from endemic areas to non-endemic areas. An enzyme-linked immunosorbent assay (ELISA) malaria antibody test has been developed that is claimed to perform better than the immunofluorescence assay test (IFAT). The assay contains antigens to both Plasmodium falciparum and Plasmodium vivax. A multicentre study was performed to evaluate the appropriateness of replacing the IFAT by the new ELISA test. MATERIAL AND METHODS: Nine French blood banks participated in this multicentre study. Two panels of samples were evaluated. The first included 4163 samples from healthy donors and was used to calculate clinical specificity of the assay. The second involved 10,995 samples, either collected retrospectively or prospectively from malaria-risk donors , was used to assess the comparative performance of the ELISA and IFAT. Discordant samples were further tested using an in-house IFAT and also tested for presence of Plasmodium DNA by polymerase chain reaction. RESULTS: The ELISA showed a clinical specificity of 99.02%. In the malaria-risk blood donors groups, the retrospective group showed a concordance rate of 92.6% (k = 0.90), while the prospective group showed a concordance rate of 97% (k = 0.46). After confirming the discordant sample results by an in-house IFAT, the k index increased to 0.81. None of the discordant samples was shown to contain Plasmodium DNA. CONCLUSION: The performance of the ELISA test in this study has confirmed its potential as a new screening test for use in blood banks, as an alternative to the IFAT in prevention of transfusion-transmitted malaria in non-endemic countries.

[Adaptation of a freezing technic with a low concentration of glycerol for preserving red blood cells resuspended in saline-adenine-glucose]. / Adaptation de la technique de congélation avec faible concentration en glycérol à des concentrés globulaires resuspendus en SAG.

We describe here a freezing method with a final low concentration of 20% glycerol and storage in a aluminium flask at - 150 degrees C. This technique allows the freezing of red blood cells concentrates collected on CPD and resuspended in SAG, without having to eliminate the SAG solution. For this we used an initial solution of 60% glycerol, which we added to the red blood cell concentrate in two steps separated by a resting time. Accordingly to the final concentration of glycerol, this freezing process is similar to the method currently used in our laboratory. This consists in one step addition V/V of 40% glycerol solution to the CPD red blood cells. The two techniques have been compared after thawing, and washing in a IBM Blood Processor 2991. The following parameters have been studied mainly: --haemoglobin level --residual glycerol --leucocytes and platelets --amount of 2-3 DPG and of intra-erythrocyte ATP. The results of the two freezing techniques are similar. They show that the SAG solution does not impair the intracellular penetration of glycerol. The in vitro qualities of the erythrocytes are well preserved.

Prevalence, severity, and risk factors of liver disease in blood donors positive in a second-generation anti-hepatitis C virus screening test.

In a cohort of 483 blood donors positive for antibody to hepatitis C virus on second-generation enzyme-linked immunosorbent, the confirmatory second-generation recombinant immunoblot assay (Ortho Diagnostic Systems) was positive in 172 cases (36%), indeterminate in 113 (23%), and negative in 198 (41%). We further studied 94 of the donors (recombinant immunoblot assay positive in 85, indeterminate in 6, and negative in 3). Alanine transaminase (ALT) activity, assayed on three occasions, was elevated in at least one assay in 85% of the 85 recombinant immunoblot assay-positive donors. Liver disease was present in 95% of these patients (chronic persistent hepatitis, 35%; chronic active hepatitis, 53%; cirrhosis, 7%). Ten of the 13 recombinant immunoblot assay-positive donors with normal ALT activity had liver disease; polymerase chain reaction testing for viral RNA was predictive of liver disease in most cases. Donors with cirrhosis differed significantly from cirrhosis-free donors in terms of age, sex ratio, ALT activity, and excessive alcohol consumption. Three of the 6 recombinant immunoblot assay-indeterminate donors (isolated C 22) who underwent histological examination had elevated ALT activity and liver disease. The 3 recombinant immunoblot assay-negative donors evaluated were free of liver disease. This study shows that anti-HCV second-generation enzyme-linked immunosorbent positivity is confirmed in fewer than 40% of blood donors by the second-generation recombinant immunoblot assay, and that liver disease is present in 95% of recombinant immunoblot assay-positive donors. Recombinant immunoblot assay positivity combined with viremia is frequently associated with the existence of liver disease, regardless of transaminase activity. Excessive alcohol consumption may be an important factor in the onset of cirrhosis in anti-HCV-positive blood donors.

Interviews of individuals diagnosed as anti-human immunodeficiency virus-positive through the screening of blood donations in the Paris area to 1994: reflections on the selection of blood donors.

BACKGROUND: One of the aims of the medical interview routinely preceding each blood donation is the identification of individuals with a risk factor for infection with the human immunodeficiency virus (HIV). STUDY DESIGN AND METHODS: Interviews were performed with individuals diagnosed as being seropositive for HIV through the systematic biologic screening of blood donations in the Paris area to establish, first, the circumstances allowing HIV-seropositive individuals to pass through the predonation medical interview and, second, the motivation of these individuals as blood donors. Risk factors of 30 HIV-infected donors identified between 1991 and 1994 were determined. RESULTS: When asked whether they recognized the eventual risk to recipients of donated blood, 14 (47%) of 30 answered positively. Fifteen (50%) admitted having given their blood to determine their HIV status. CONCLUSION: These individuals did not exclude themselves from blood donation and probably hid their risk factor(s) at the predonation interview in order to be accepted as blood donors.

Risk factors for hepatitis C virus infection in hepatitis C virus antibody ELISA-positive blood donors according to RIBA-2 status: a case-control survey.

Studies of blood donors positive for antibody to hepatitis C virus on enzyme-linked immunosorbent assay are probably biased by the large number of false-positive results. We evaluated the epidemiological and biological characteristics of 177 such donors with regard to the confirmatory second-generation RIBA test (Ortho Diagnostic Systems) and have compared the results to those from an age- and sex-matched control group of 177 donors negative for antibody to hepatitis C virus on enzyme-linked immunosorbent assay. Second-generation recombinant immunoblot assay was positive in 38% of cases, indeterminate in 6% and negative in 56%. The case-control study showed a significantly higher frequency of intravenous drug abuse (27% vs. 0%), blood transfusion (22% vs. 9%), history of jaundice (21% vs. 7%), elevated ALT level (49% vs. 4%) and HBc antibody positivity (32% vs. 7%) in second-generation recombinant immunoblot assay-positive donors. No such differences were found between the second-generation recombinant immunoblot assay-negative donors and their controls. The 35 second-generation recombinant immunoblot assay-positive donors without histories of transfusion or intravenous drug abuse had a significantly higher frequency of surgery with major blood loss or prolonged stays in areas of hepatitis B virus endemicity than did their controls (20% vs. 0% and 49% vs. 26%, respectively). In conclusion, at least one risk factor for HCV infection was identified in 82% of the second-generation recombinant immunoblot assay-positive donors, 91% of whom could have been identified on the basis of these risk factors, ALT level and presence of HBc antibody.

Interviews with anti-HIV-positive individuals detected through the systematic screening of blood donations: consequences on predonation medical interview.

This study is based upon interviews with 74 individuals found to be human immunodeficiency virus (HIV)-seropositive through the screening of blood donations between January 1988 and December 1990. The donation history and the risk factor of HIV infection were established. Questions about the use of blood donation as a diagnostic test and on the notion of a predonation medical interview evoking the risk factor were asked. The majority of the individuals had a risk factor of HIV infection and had given their blood for serological testing. This data can help to adapt the predonation medical interview to the present epidemiological context of HIV infection. The improvement of this interview will contribute to the decrease of the residual transfusional risk of HIV infection.

Epidemiologic comparison of human T-lymphotropic virus type I-infected blood donors from endemic and nonendemic regions over a 3-year period.

BACKGROUND: Screening for human T-lymphotropic virus type I (HTLV-I) infection became systematic in 1989 in the French West Indies for blood from all donors and in France for blood from natives of endemic areas; in 1990, it was extended to blood from donors with at-risk sex partners and in July 1991 to blood from all donors. STUDY DESIGN AND METHODS: The epidemiologic characteristics of individuals found through the screening of donated blood to be HTLV-I infected were compared for an endemic region (Guadeloupe, French West Indies) and a nonendemic region (Paris area) over a 3-year period (1989 through 1991). RESULTS: In Guadeloupe, 131 HTLV-I-infected individuals were detected in the screening of 28,801 units; in the Paris area, 38 HTLV-I-infected donors were detected in the screening of 109,824 units. All Guadeloupean HTLV-I-infected donors were natives of endemic areas. Among the 38 Parisian HTLV-I-infected donors, 21 were natives of endemic areas, 10 were natives of endemic areas and had received transfusions, 2 were whites who had received transfusions, and 5 were whites who had had heterosexual contact with natives of endemic areas. The percentage of HTLV-I-infected individuals whose blood would have been excluded because of positivity for one or more markers for other viruses did not significantly change over the study period and did not significantly differ between regions (41%). Among the eight Parisian HTLV-I-infected blood donors detected after July 1991, six would not have been detected without the biologic screening. CONCLUSION: The generalization of biologic screening of HTLV-I-infected donated blood in France was useful for the prevention of HTLV-I and HTLV type II infections through transfusion.

[Technical, immunological and clinical study of the performance of blood filtration using Erypur filters]. / Etude technique, immunologique et clinique des performances de filtration du sang à l'aide de l'appareil Erypur.

Technical, immunological and clinical parameters of blood filtration through Erypur filters were evaluated. Red cell concentrates from which the buffy coat had not been eliminated were used throughout the study. Filtration procedure is fully automated, simple and takes about 15 minutes. 98% of leukocytes and 96% of platelets were found to be retained by the filter. Residual leukocytes were less than 3.10(7) in 68% and residual platelets less than 1.10(10) in 93% of filtered units. The incidence of lymphocyte alloantibody formation was 14.9% in the group of 67 patients who received 210 filtered units and 27.9% in the control group of 93 patients who were transfused with 298 non-filtered units. 330 filtered units were transfused to 107 carefully monitored patients and caused no reaction. 40 of these patients had polyspecific HLA antibodies and/or a documented history of non-hemolytic transfusion reactions. The clinical usefulness of Erypur filtered blood in the prevention of such reactions may thus be confirmed.

A comparative evaluation of the sensitivity of five anti-hepatitis C virus immunoblot assays.

BACKGROUND AND OBJECTIVES: In order to compare sensitivity, five anti-hepatitis C virus (HCV) immunoblot assays were tested (Deciscan plus, Inno-Lia III, Matrix, Murex Western blot and RIBA-3). MATERIALS AND METHODS: The test panel (50 samples for each assay) included 6 anti-HCV-negative samples and 44 samples from 36 HCV-infected subjects. RESULTS: There were minor differences in core reactivity among the assays. The smallest number of NS3-reactive results occurred with the Murex and Matrix assays, and the smallest number of NS4 reactives with RIBA-3 and Matrix. Among the 20 discrepant results for NS5 there was one clear false-negative with Inno-Lia. Only 28 of the 50 samples of the panel gave the same results in all the assays: 5 negatives and 23 positives. One of the 6 negative samples were indeterminate in 3 assays. Eighteen of the 21 other divergent results were interpreted as either indeterminate or positive, a common reactivity being exhibited by all 5 assays. The most important discrepancies occurred on 3 HCV-RNA-positive samples which came up negative in some assays: 2 samples with isolated NS3 reactivity were negative by Matrix and Murex Western blot, 1 of them being also negative by Inno-Lia III; another sample was negative by RIBA-3 and Matrix due to weak signals (< 1) on core and NS3 proteins, which did not exceed 1+ with the other assays. CONCLUSIONS: With more uniform criteria for interpretation, the results would have been less divergent. Some assays should improve their sensitivity to the NS3 protein.

GB virus type C/HGV markers in HCV RNA-positive French blood donors: correlation with HCV genotypes and risk factors.

BACKGROUND: Exposure to GB virus type C/HGV (GBV-C/HGV) could be determined by detection either of RNA by RT-PCR or of antibodies of the envelope protein E2. STUDY DESIGN AND METHODS: The aim of the study was to determine the proportion of the GBV-C/HGV markers of infection in a blood donor population infected with HCV and to identify GBV-C/HGV routes of transmission that are associated with HCV genotypes and risk factors. RESULTS: Among 306 HCV RNA-positive blood donors, the proportion of GBV-C/HGV RNA-positive donors and anti-E2-positive donors was 19.3 percent (95% CI = 15.0-24.2%) and 42.1 percent (95% CI = 36.6-47.9%), respectively. Exposure to GBV-C/HGV (RNA or anti-E2) was significantly associated with the risk factor of IV drug use. There was a trend toward association with HCV subtypes 1a and 3a, probably because these HCV subtypes are the most frequent in IV drug users. No correlation was observed between ALT elevation and the presence of GBV-C/HGV RNA. CONCLUSION: In persons with HCV infection, IV drug use seems to be a major route of GBV-C/HGV transmission. Precautions taken to avoid HCV infection will probably also decrease GBV-C/HGV transmission.

[Practical approach to the discovery of serum markers for infection by hepatitis B virus (HBs antigen or anti HBc antibody). Hypertransaminasemia or the two anomalies in a blood donor]. / Attitude pratique devant la découverte de marqueurs sériques d'infection par le virus de l'hépatite B (antigène HBs et/ou anticorps anti-HBc). Une hypertransaminasémie ou les deux anomalies chez un donneur de sang.

By now in France HBV seric markers (Ag HBs and Ac HBc) and transaminases level (ALT) screenings are compulsory by law in blood donors. People whose blood donation is discarded should be informed. A clinical, epidemiological and virological survey of such donors is required to differentiate healthy Ag HBs carriers and patients suffering from hepatitis B (who may eventually be treated). Similar guidelines may be recommended for the blood donors presenting high transaminases level without HBV seric markers in order to find a cause for such impaired biochemical tests: overweight, alcohol, drug consumption, auto-immune liver disease, genetic disorder, Non-A, Non-B, Non-C hepatitis....

[Analytical study of the Abbott and Ortho tests for screening and confirmation of anti-HCV antibodies. Le groupe de Travail "Hépatites virales" de la Société Française de Transfusion Sanguine]. / Etude analytique des trousses de dépistage et de confirmation des anticorps anti-VHC Abbott et Ortho.Le groupe de Travail "Hépatites Virales" de la Société Française de Transfusion Sanguine.

Anti-HCV systematic screening on blood donation was mandatory in France since first of March 1991. Two laboratories (Ortho-Chiron and Abbott) have introduced in Europe successively two kinds of hepatitis C positive diagnosis with 1st and 2nd generation ELISA screening and confirmatory assays. The aim of this multicentric study was to evaluated the sensibility and specificity of these tests. For that, they used 10,090 blood sera. As a result we have seen that the new "second generation" screening assays have a higher sensitivity without less of specificity for the confirmatory tests.

[Hepatitis C virus and transfusion: policy regarding the donation and the donor. Groupe "Hépatites Virales" de la Société Nationale de Transfusion Sanguine]. / Virus de l'hépatite C et transfusion: attitude vis-à-vis du don et du donneur. Groupe "Hépatites Virales" de la Société Nationale de Transfusion Sanguine.

Hepatitis C virus (HCV) discovery and introduction of anti-HCV antibodies screening in blood transfusion imply the necessity of a good blood donations and blood donors policy. Detection of a seropositivity during the screening must be completed with a confirmatory test. The results are directly used to inform donors and define the blood products policy. Donors with positive results on confirmatory test are discarded and have physical and biological examinations in hepatology. Individuals with indeterminate or negative results must be retested for the HCV serology. Furthermore, because of a rapid improvement in the fields of technology, diagnosis and therapy of HCV, an adaptation of the policy is necessary.