Assessment of Liaison XL Murex Chagas diagnostic performance in blood screening for Chagas disease using a reference array of chimeric antigens.

BACKGROUND: Chagas disease (CD) serological screening at blood banks is usually performed by a single highly sensitive serological assay, with chemiluminescent immunoassays (CLIAs) being the method of choice. CLIAs employ recombinant, fusion peptides and/or chimeric antigens that selectively capture anti-Trypanosoma cruzi antibodies. However, despite high sensitivity, the ability of these tests to identify CD-positive cases should be evaluated against T. cruzi strains circulating in specific locales. Herein, we used a latent class analysis (LCA) approach employing an array of four chimeric antigens to assess the diagnostic performance of the Liaison XL Murex Chagas CLIA for the detection of anti-T. cruzi IgG in serum samples. STUDY DESIGN AND METHODS: The study included a panel of 5014 serum samples collected from volunteer blood donors at the Hematology and Hemotherapy Foundation of the State of Bahia, submitted to anti-T. cruzi antibody detection using Liaison Chagas CLIA and LCA as a reference test in the absence of a gold standard. RESULTS: LCA classified 4993 samples as negative, while positivity for T. cruzi antibodies was predicted in 21 samples. Compared with LCA, CLIA demonstrated sensitivity and specificity of 76.2% and 99.5%, respectively, providing an overall accuracy of 99.4%. DISCUSSION: In blood banks lacking a de facto highly sensitive screening immunoassay, the low sensitivity offered by Liaison Chagas CLIA renders it unsuitable for standalone use in serological screening procedures for CD. Moreover, blood banks are encouraged to carefully assess the ability of diagnostic methods to identify local T. cruzi strains in circulation.

Dried Blood Spot Technique-Based Liquid Chromatography-Tandem Mass Spectrometry Method as a Simple Alternative for Benznidazole Pharmacokinetic Assessment.

Chagas disease (CD) is recognized as one of the major neglected global tropical diseases. Benznidazole (BNZ) is the drug of choice for the treatment of adults, young infants, and newborns with CD. However, the pharmacokinetics (PK) of BNZ have been poorly evaluated in all age groups, with consequent gaps in knowledge about PK-pharmacodynamic relationships in CD. The purpose of this study was to develop and validate a bioanalytical method to quantify BNZ levels in small-volume whole-blood samples collected as dried blood spots (DBS). The analysis was performed using high-performance liquid chromatography-positive electrospray tandem mass spectrometry. PK evaluation in healthy male volunteers was conducted to verify the correlation between DBS and plasma BNZ concentrations. The calibration curve was linear from 50 to 20,000 ng · ml-1 Intra- and interday precision and bias values were less than 14.87% (n = 9) and 9.81% (n = 27), respectively. The recovery rates ranged from 94 to 100% with no matrix effect. There was no hematocrit level effect in a range of 20 to 70%. The PK results obtained from DBS and plasma were comparable (r2 = 0.8295) and equivalent to previously published information on BNZ. BNZ in DBS was stable at room temperature for more than one year. This article describes the first microsampling method for measuring BNZ levels in DBS that has the potential to facilitate broad implementation of PK in clinical trials involving adult and pediatric patients in remote areas and helps to address existing knowledge gaps in the treatment of CD.

Cost-Effectiveness of Blood Donation Screening for Trypanosoma cruzi in Mexico.

An estimated 2 million inhabitants are infected with Chagas disease in Mexico, with highest prevalence coinciding with highest demographic density in the southern half of the country. After vector-borne transmission, Trypanosoma cruzi is principally transmitted to humans via blood transfusion. Despite initiation of serological screening of blood donations or donors for T. cruzi since 1990 in most Latin American countries, Mexico only finally included mandatory serological screening nationwide in official Norms in 2012. Most recent regulatory changes and segmented blood services in Mexico may affect compliance of mandatory screening guidelines. The objective of this study was to calculate the incremental cost-effectiveness ratio for total compliance of current guidelines from both Mexican primary healthcare and regular salaried worker health service institutions: the Secretary of Health and the Mexican Institute for Social Security. We developed a bi-modular model to analyze compliance using a decision tree for the most common screening algorithms for each health institution, and a Markov transition model for the natural history of illness and care. The incremental cost effectiveness ratio based on life-years gained is US$ 383 for the Secretary of Health, while the cost for an additional life-year gained is US$ 463 for the Social Security Institute. The results of the present study suggest that due to incomplete compliance of Mexico's national legislation during 2013 and 2014, the MoH has failed to confirm 15,162 T. cruzi infections, has not prevented 2,347 avoidable infections, and has lost 333,483 life-years. Although there is a vast difference in T. cruzi prevalence between Bolivia and Mexico, Bolivia established mandatory blood screening for T.cruzi in 1996 and until 2002 detected and discarded 11,489 T. cruzi -infected blood units and prevented 2,879 potential infections with their transfusion blood screening program. In the first two years of Mexico's mandated program, the two primary institutions failed to prevent due to incomplete compliance more potential infections than those gained from the first five years of Bolivia's program. Full regulatory compliance should be clearly understood as mandatory for the sake of blood security, and its monitoring and analysis in Mexico should be part of the health authority's responsibility.

Chagas disease and transfusion medicine: a perspective from non-endemic countries.

In the last decades, increasing international migration and travel from Latin America to Europe have favoured the emergence of tropical diseases outside their "historical" boundaries. Chagas disease, a zoonosis endemic in rural areas of Central and South America represents a clear example of this phenomenon. In the absence of the vector, one of the potential modes of transmission of Chagas disease in non-endemic regions is through blood and blood products. As most patients with Chagas disease are asymptomatic and unaware of their condition, in case of blood donation they can inadvertently represent a serious threat to the safety of the blood supply in non-endemic areas. Since the first cases of transfusion-transmitted Chagas disease were described in the last years, non-endemic countries began to develop ad hoc strategies to prevent and control the spread of the infection. United States, Spain, United Kingdom and France first recognised the need for Trypanosoma cruzi screening in at-risk blood donors. In this review, we trace an up-to-date perspective on Chagas disease, describing its peculiar features, from epidemiological, pathological, clinical and diagnostic points of view. Moreover, we describe the possible transmission of Chagas disease through blood or blood products and the current strategies for its control, focusing on non-endemic areas.

Concordancia de dos pruebas serológicas para el diagnóstico de la enfermedad de Chagas / Concordance of two serological tests for the diagnosis of Chagas disease

Objetivo Establecer la concordancia entre un Ensayo Inmunoenzimático Ligado a una Enima Casera (ELISA) y la Inmunofluorescencia Indirecta (IFI) para el diagnóstico de infección por T. cruzi empleando eluidos sanguíneos. Metodología Se realizó un estudio de evaluación de tecnología diagnóstica y muestreo de corte transversal a 650 habitantes de una zona endémica de Colombia. Se determinó el área bajo la curva de operador-receptor (del inglés ROC) y se usó la IFI estandarizada en eluidos sanguíneos como gold standard. Se estableció el punto de corte para el ELISA, así como la concordancia entre las lecturas. Resultados El ELISA presentó una concordancia de 0,99 (IC95 %: 0,989-0,992) entre las lecturas realizadas y una curva ROC de 0,9795. El punto de corte establecido fue 0,5 de absorbancia en la prueba de ELISA. 16,6 % fueron positivas para anticuerpos anti-T. cruzi por ELISA y 10,9 % por IFI. Conclusiones El ELISA mostró buena concordancia frente a IFI, por lo tanto es una buena elección diagnóstica para la población que vive en áreas remotas.

Objetive Establish the concordance between in-house ELISA and IIF for the diagnosis of infection with T. cruzi using blood eluates. Methodology A study of diagnostic technology evaluation and cross-sectional sample of 650 residents of an endemic area of Colombia was conducted. It was determined the Receiver Operating Characteristic curve (ROC) and IIF was used as a gold standard. It was established the cutoff for the ELISA and the correlation between readings. Results The in-house ELISA it was an agreement of 0.99 (95 % CI: 0.989 to 0.992) between the two readings taken and the area for the ROC curve was 0.9795. The cutoff was set at 0.5 absorbance in the ELISA test. 16.6 % were positive by ELISA and 10.9 % by IIF. Conclusions The in-house ELISA showed good concordance compared to the IIF, so it is a good choice diagnostic for the population living in remote areas.

Applicability of an optimized non-conventional flow cytometry method to detect anti-Trypanosoma cruzi immunoglobulin G for the serological diagnosis and cure assessment following chemotherapeutic treatment of Chagas disease.

One of the challenges on immunodiagnostic of Chagas disease in endemic areas has been the search for more practical and safe antigenic preparation that provides tests with higher sensitivity and specificity, with low cross-reactivity. A new approach using fixed Trypanosoma cruzi epimastigotes to detect IgG reactivity was investigated previously. In order to continue this investigation, this study aimed at optimizing the flow cytometry-based method to the diagnosis of Chagas disease patients after specific chemotherapy. To achieve our goal, serum samples from 93 subjects - 52 adults chronically infected by T. cruzi, and 41 uninfected controls were tested by flow cytometry. Secondly, serum samples from patients Treated Cured and Treated Uncured from Chagas disease were also tested to evaluate the potential of the method on assessing cure. After establishing the ideal serum dilution and cut off, 121 serum samples from patients with other endemic infections were tested to check cross-reactivity. The results showed that parasite staining with Evan's blue dye eliminated debris, allowing trustworthy analysis of anti-fixed epimastigote IgG reactivity. The applicability of the method to diagnose Chagas disease was confirmed by the high sensitivity (98.1%) and specificity (100%) found. This method also contributed for post-therapeutic assessment of cure, identifying 94.1% of Treated Uncured and 83.3% of Treated Cured patients. Cross-reactivity was observed in a very low number (6.7%). On the whole, these data highly recommend the use of anti-fixed T. cruzi epimastigote IgG reactivity by flow cytometry to the diagnosis and cure monitoring of Chagas disease in endemic areas.

An inexpensive antigen for serodiagnosis of Chagas' disease.

UNLABELLED: In this prospective study we evaluated the performance characteristics of a specific and sensitive antigen preparation (AgA) used in an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-Trypanosoma cruzi antibodies in serum samples, for Chagas' disease diagnosis. The antigen production was achieved by combination of nutritional stress and autoclaving the parasites. Specificity and sensitivity were evaluated in two separate tests, using 152 sera from healthy individuals and 175 sera from Chagas' patients (70 by xenodiagnosis). Cross-reactivity was tested using 289 sera from patients who had a parasitological diagnosis of a disease known to induce antigenic responses towards T. cruzi. All of these sera were tested with our AgA-ELISA and with 3 commercial diagnosis kits. To evaluate the agreement of results between our AgA-ELISA and a "gold standard" test for Chagas, we tested 566 sera from an endemic area. RESULTS: sensitivity and specificity were 100%; cross-reactivity was the lowest compared with commercial kits. Overall agreement with the gold standard test was excellent (kappa = 0.92). AgA-ELISA exhibits levels of sensitivity, specificity and cross-reactivity comparable or superior to those shown, obtained with the commercial kits used in our country, while being at least 10 times less expensive. This balance between diagnostic accuracy and cost makes AgA-ELISA useful for blood bank screening in poor regions of the world suffering from Chagas' disease. Further validations of this antigenic formulation in other countries are necessary.

Knowledge deficits regarding Chagas disease may place Mexico's blood supply at risk.

Prevention of transfusion-related Chagas disease in Mexico City depends on targeted questionnaire-based screening of donors by nurses at blood banks. To assess potential problems with this strategy, surveys were distributed to the nurses who screen donors in a random sampling of nine blood banks in Mexico City, to measure appropriate knowledge about Chagas disease. We found that 80% (95% CI 68-92%) of nurses answered at least one of the three donor risk factor questions incorrectly, which may fail to trigger confirmatory laboratory testing of potentially infected units. If this knowledge deficit is widespread, up to 680,000 units (95% CI 578,000-782,000 units) of donated blood could be potentially contaminated with Chagas disease in Mexico. In place of targeted screening, routine laboratory testing of all donated blood would be a cost-effective method to safeguard blood recipients from iatrogenic Chagas disease.

Update on seroprevalence of anti-Trypanosoma cruzi antibodies among blood donors in northeast Mexico.

Chagas disease has become frequent in non-endemic areas, where it can be transmitted by blood transmission. Therefore, we explored seroprevalence of anti-Trypanosoma cruzi antibodies among blood donors at the Cardiology Hospital, Mexican Institute of Social Security at Monterrey, Nuevo León, by both an enzyme-linked immunosorbent assay and indirect hemagglutination. Blood samples from 1,000 healthy blood donors were selected. A seropositivity of 2.8% was shown among the studied population, of which 2.59% (21/809) were inhabitants of Nuevo León, whereas 3.07% (2/65) and 3.96% (5/126) were from Coahuila and Tamaulipas, respectively. Our result is higher than that of a previous study from 1998, where a prevalence of 0.5% was reported. This once again corroborates the importance of installing a surveillance program to detect and prevent the transfusion of T. cruzi from asymptomatic blood donors in blood banks located in urban cities recognized as non-endemic.

Cost-effectiveness of implementation methods for ELISA serology testing of Trypanosoma cruzi in California blood banks.

The first U.S. ELISA test for T. cruzi antibodies was licensed by the Food and Drug Administration (FDA) on December 13, 2006. Blood banks have begun screening in absence of FDA recommendations for best implementation methods. We surveyed 2,029 blood donors at five California sites with three risk-based Chagas risk-screening questions. Semi-Markov models compared the cost-effectiveness of three testing strategies. 30% of donors screened positively. Screening all dominated doing nothing, being less costly, and saving more lives. The choice to "screen and test" compared with "testing all" varied by Chagas prevalence, "screening and testing" being cost-effective for high (0.004) and low (0.00004) prevalences, and "testing all" cost-effective for moderate risk (0.0004). It is cost-effective to screen by ELISA rather than do nothing. The best strategy depends on site-specific risk. Census estimates of Hispanics do not predict donor risk well. We suggest using our screening questions to determine risk level and most cost-effective testing strategy.

Assessment of a travel question to identify donors with risk of Trypanosoma cruzi: operational validity and field testing.

BACKGROUND: Because Trypanosoma cruzi (T. cruzi) infection in Canada and the United States is largely contracted in endemic countries, targeted testing of blood donors with risk travel may improve safety. The operational validity of a travel question suitable for donor screening was tested, and it was field-tested. STUDY DESIGN AND METHODS: After 1331 donors completed a short travel question, operational validity was assessed by detailed travel histories in face-to-face interviews. Two nationwide donor surveys were carried out assessing donor responses to similar travel questions in 2001 (13,623 donors) and in 2006 (20,037 donors). All donors in Toronto, Ontario, answered a travel question in 1997 and those born in or who spent 6 months or more in Mexico, Central America, or South America were tested for antibody to T. cruzi. RESULTS: There was 97.3 percent agreement between the travel question and detailed interviews, with 15 donors (1.1%) failing to acknowledge risk travel (false-negative questioning responses). Of these, 6 donors were born there and 7 others had less than 1 year of cumulative travel. In 2001 and 2006, there were 2.1 and 2.0 percent of donors with risk travel, respectively, but 16.5 and 11.2 percent of these donors were identified only because they were born there (travel not acknowledge). There were 1337 (1.6%) donors in Toronto in 1997 with risk travel and none were positive for the presence of T. cruzi antibody. CONCLUSION: Donors can answer a short question about cumulative time in Latin America with similar accuracy to detailed questioning, but screening questions should also include country of birth.

[Prevalence of antibodies against Trypanosoma cruzi in blood bank donors from the IMSS General Hospital in Orizaba, Veracruz, Mexico]. / Prevalencia de anticuerpos contra Trypanosoma cruzi en donadores de sangre del IMSS, Orizaba, Veracruz, México.

OBJECTIVE: To estimate the prevalence of antibodies against Trypanosoma cruzi in blood donors from Hospital General Regional (HGRO) of the Mexican Institute of Social Security (IMSS per its abbreviation in Spanish). MATERIAL AND METHODS: Between October 2001 and January 2002, blood samples were collected from voluntary donors at the blood bank of the Hospital General Regional of IMSS in Orizaba; Veracruz, Mexico. The samples were assayed for anti-T. cruzi by ELISA, Western blot and IFI, using a recombinant protein (MBP::Hsp70), and crude extract from epimastigotes. RESULTS: A total of 420 blood donors were studied; two of them were seropositive for ELISA,Western blot and IFI, with a seroprevalence of 0.48%. CONCLUSIONS: Some blood donors at the HGRO hospital were seropositive for T. cruzi, showing the risk of contamination by blood transfusion. Routine serologic screening with highly sensitive and specific immunological techniques are needed.

[Chronic Chagas' disease: from xenodiagnosis and hemoculture to polymerase chain reaction]. / Doença de Chagas crônica: do xenodiagnóstico e hemocultura à reação em cadeia da polimerase.

Although there has been an improvement in the diagnosis of chronic Chagas' disease, the low sensitivity of indirect parasitological tests is a drawback to its application in diagnosis and post-therapeutic control. Polymerase chain reaction (PCR) has limited use in routine diagnosis due to the need of specific laboratory facilities, common DNA cross-contamination, and high costs. At the same time, the high variability of PCR results found in different regions of Brazil raises some questions concerning its applicability for diagnosis. PCR's high specificity is indicative that it can be used as a confirmation method in inconclusive serology diagnosis as well as an auxiliary method in pos-therapeutic control of chronic Chagas' disease when comparing to serology and parasitological techniques. It is discussed here the applicability of molecular and indirect parasitological methods in the diagnosis and post-therapeutic control of chronic Chagas' disease based on the literature published from 1954 to 2001.

Transfusion-transmitted infectious diseases in Argentina, 1995 through 1997.

BACKGROUND: Assessment of the safety of the blood supply, the quality of screening procedures, and the risk of transfusion transmission of infectious diseases in any country can be estimated by reviewing the records of blood donations and screening procedures and the prevalence of serologic markers of infectious diseases. STUDY DESIGN AND METHODS: Information on blood donors, particularly the number of screened donors, and on the prevalence of serologic markers of infectious diseases was available from Argentina for 1995 through 1997. This information permitted the estimation of the risks and costs of preventing transfusion transmission of infectious diseases within the country during this period. RESULTS: Screening coverage was higher in the private sector. The proportion of donors screened for HIV increased from 84.52 percent in 1995 to 97.97 percent in 1997; in the same period, serologic screening for HbsAg increased from 83.71 percent to 98.48 percent; that for HCV from 69. 92 percent to 97.83 percent; and that for syphilis from 87.94 percent to 98.71 percent. One hundred percent of donors were screened for Trypanosoma cruzi throughout the period. The overall prevalence of HIV per year varied from 2.42 to 3.36 per 1,000 donors; that of HBV, from 5.80 to 9.76 per 1,000; of HCV, from 7.39 to 16.61 per 1,000; and of syphilis, from 5.25 to 7.65 per 1,000. The overall prevalence of antibodies to T. cruzi ranged from 36.53 to 49.20 per 1,000 donors. The overall index of the spread of infectious viral disease through blood transfusion decreased from 47. 74 per 10,000 donations in 1995 to 4.75 per 10,000 in 1997. The ratio of acquired infections to donations improved from 1:209 to 1:2, 102 during the same period. The risk of T. cruzi infection from 1995 through 1997 was, in theory, nil, given the 100-percent screening. The greatest threat to the quality of the blood supply throughout the period studied was HCV. CONCLUSION: The status of the blood supply in Argentina improved steadily from 1995 to 1997, as shown by the increase in screening coverage.

[The assessment of parasitemia in women who are carriers of Trypanosoma cruzi infection during and after pregnancy]. / Avaliação da parasitemia em mulheres portadoras de infecção pelo Trypanosoma cruzi durante e após a gestação.

This paper presents an evaluation of the parasitemic profiles of 119 women chronically infected with T. cruzi. Xenodiagnosis (xenos) were applied during (465 xenos) and after pregnancy (363 xenos) in order to detect possible variations in parasitemia in these periods. The frequency of positive xenos was greater during than after gestation. Otherwise, the frequency of infected triatomines was wore elevated during pregnancy, indicating higher parasitemic levels in this period. Only 17% of the studied women had two or more positive xenos during pregnancy. In these mothers the difference between the frequencies of positive xenos during and after gestation was high, suggesting the occurrence of exacerbation of infection at least in some women.