Towards a New Strategy for Diagnosis of Congenital Trypanosoma cruzi Infection

Abràs, Alba; Muñoz, Carmen; Ballart, Cristina; Berenguer, Pere; Llovet, Teresa; Herrero, Mercedes; Tebar, Sílvia; Pinazo, María Jesús; Posada, Elizabeth; Marti, Carmen Gonzalez; Fumadó, Victòria; Bosch, Jordi; Coll, O.; Juncosa, T; Ginovart, Gemma; Armengol, Josep; Gascón, Joaquím; Portús, Montserrat; Gállego, Montserrat

doi:10.1128/jcm.02248-16

Cited by 23 publications

(21 citation statements)

References 59 publications

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“…Because of the lack of a single reference standard test, the possibility of cross-reactivity, and the biological diversity and genetic polymorphism of T. cruzi , which is due to the existence of six discrete typing units (DTUs) of the parasite in different geographical areas [ 10 ] , the WHO recommends that a diagnosis of chronic CD be based on two positive results obtained using two different methods: a conventional test followed by a non-conventional assay [ 9 ]. Serological diagnosis remains a challenge and screening schemes have been implemented only recently, mainly for blood and organ donors, pregnant women, and newborns [ 11 – 13 ], in an effort to control the transmission of CD [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Serological evaluation for Chagas disease in migrants from Latin American countries resident in Rome, Italy

et al. 2018

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BackgroundChagas disease (CD) is a systemic parasitic infection caused by the protozoan Trypanosoma cruzi, whose chronic phase may lead to cardiac and intestinal disorders. Endemic in Latin America where it is transmitted mainly by vectors, large-scale migrations to other countries have turned CD into a global health problem because of its alternative transmission routes through blood transfusion, tissue transplantation, or congenital. Aim of this study was to compare the performance of two commercially available tests for serological diagnosis of CD in a group of Latin American migrants living in a non-endemic setting (Rome, Italy). The study was based on a cross-sectional analysis of seroprevalence in this group. Epidemiological risk factors associated to CD were also evaluated in this study population.MethodsThe present study was conducted on 368 subjects from the Latin American community resident in Rome. Following WHO guidelines, we employed a diagnostic strategy based on two tests to detect IgG antibodies against T. cruzi in the blood (a lysate antigen-based ELISA and a chemiluminescent microparticle CMIA composed of multiple recombinant antigens), followed by a third test (an immunochromatographic assay) on discordant samples.ResultsOur diagnostic approach produced 319/368 (86.7%) concordant negative and 30/368 (8.1%) concordant positive results after the first screening. Discrepancies were obtained for 19/368 (5.2%) samples that were tested using the third assay, obtaining 2 more positive and 17 negative results. The final count of positive samples was 32/368 (8.7% of the tested population). Increasing age, birth in Bolivia, and previous residence in a mud house were independent factors associated with T. cruzi positive serology.ConclusionsSerological diagnosis of CD is still challenging, because of the lack of a reference standard serological assay for diagnosis. Our results reaffirm the importance of performing CD screening in non-endemic countries; employing a fully automated and highly sensitive CMIA assay first could be a cost- and resource-effective strategy for mass screening of low-risk patients. However, our results also suggest that the WHO strategy of using two different serological assays, combined with epidemiological information, remains the best approach for patients coming from endemic countries.

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Section: Introductionmentioning

confidence: 99%

Serological evaluation for Chagas disease in migrants from Latin American countries resident in Rome, Italy

et al. 2018

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“…The use of light microscopy to detect parasitic forms of T. cruzi is recommended in cases in which there are clinical or epidemiological suspicions of an acute phase, since a greater amount of trypomastigotes circulating in blood can be observed during the first weeks after infection (Vega and Náquira, 2006;Abras et al, 2017). The puppy found to be positive using light microscopy and PCR in the transversal study, in which antibodies against the agent or clinical signs were not found, indicates that the puppy had been recently infected and was in an acute phase of infection.…”

Section: Discussionmentioning

confidence: 99%

“…The finding of a PCR-positive dog in May 2016 in the longitudinal study with enlarged popliteal lymph nodes and which was found in the following examination to be PCR-negative but serologically positive, indicates that the dog was possibly ending the acute phase in May, and by the time of the next examination had entered to the indeterminate phase. This phase is characterized by a reduction of parasitemia due to intracellular infection of cardiac cells, where the parasite continues replicating itself for many years without causing evident clinical signs (Vega and Náquira, 2006;Abras et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

Canine trypanosomiasis in an endemic Costa Rican community: Demonstration of the active infection cycle

Bonilla

Castro-Vásquez

Herrero-Acosta

et al. 2019

Veterinary Parasitology: Regional Studies and Reports

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A cross-sectional study was conducted to determine the prevalence of canine trypanosomiasis in an endemic community of Costa Rica. The indirect hemagglutination and indirect immunofluorescence assay yielded positive results in 6.4% (20/314) of canine samples analyzed; polymerase chain reaction (PCR) and light microscopy yielded positive results in one dog. Subsequently, a longitudinal study was carried out with 55 negative T. cruzi canines in the cross-sectional study. These dogs were divided into two groups: Group 1, which consisted of 25 individuals that lived in dwellings where triatomines were found in their homes; and Group 2, which consisted of 30 dogs that lived in dwellings where triatomines were not found during the previous study in their homes. Seroconversion occurred in six dogs (10.9%) in Group 1 in the first months of the year (dry season); these dogs remained seropositive until the end of the study. Only one of the six seropositive canines was also found positive once in T. cruzi PCR. The analysis of the amplified T. cruzi sequences of dogs and triatomines showed that all of them belonged to the TcI lineage. It is recommended that residents be made aware of the need to eliminate vectors in their homes and their surroundings.

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“…Nevertheless, serological diagnosis has certain disadvantages: the persistence of positive results in chronically infected patients for years after treatment [ 20 ], the possibility of cross-reactions with other trypanosomatids like Trypanosoma rangeli or Leishmania spp. [ 21 ], and the transmission of passive antibodies from mother to newborn [ 22 – 24 ].…”

Section: Introductionmentioning

confidence: 99%

Introducing automation to the molecular diagnosis of Trypanosoma cruzi infection: A comparative study of sample treatments, DNA extraction methods and real-time PCR assays

et al. 2018

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BackgroundPolymerase chain reaction (PCR) has become a useful tool for the diagnosis of Trypanosoma cruzi infection. The development of automated DNA extraction methodologies and PCR systems is an important step toward the standardization of protocols in routine diagnosis. To date, there are only two commercially available Real-Time PCR assays for the routine laboratory detection of T. cruzi DNA in clinical samples: TCRUZIDNA.CE (Diagnostic Bioprobes Srl) and RealCycler CHAG (Progenie Molecular). Our aim was to evaluate the RealCycler CHAG assay taking into account the whole process.Methodology/Principal findingsWe assessed the usefulness of an automated DNA extraction system based on magnetic particles (EZ1 Virus Mini Kit v2.0, Qiagen) combined with a commercially available Real-Time PCR assay targeting satellite DNA (SatDNA) of T. cruzi (RealCycler CHAG), a methodology used for routine diagnosis in our hospital. It was compared with a well-known strategy combining a commercial DNA isolation kit based on silica columns (High Pure PCR Template Preparation Kit, Roche Diagnostics) with an in-house Real-Time PCR targeting SatDNA. The results of the two methodologies were in almost perfect agreement, indicating they can be used interchangeably. However, when variations in protocol factors were applied (sample treatment, extraction method and Real-Time PCR), the results were less convincing. A comprehensive fine-tuning of the whole procedure is the key to successful results. Guanidine EDTA-blood (GEB) samples are not suitable for DNA extraction based on magnetic particles due to inhibition, at least when samples are not processed immediately.Conclusions/SignificanceThis is the first study to evaluate the RealCycler CHAG assay taking into account the overall process, including three variables (sample treatment, extraction method and Real-Time PCR). Our findings may contribute to the harmonization of protocols between laboratories and to a wider application of Real-Time PCR in molecular diagnostic laboratories associated with health centers.

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Towards a New Strategy for Diagnosis of Congenital Trypanosoma cruzi Infection

Cited by 23 publications

References 59 publications

Serological evaluation for Chagas disease in migrants from Latin American countries resident in Rome, Italy

Serological evaluation for Chagas disease in migrants from Latin American countries resident in Rome, Italy

Canine trypanosomiasis in an endemic Costa Rican community: Demonstration of the active infection cycle

Introducing automation to the molecular diagnosis of Trypanosoma cruzi infection: A comparative study of sample treatments, DNA extraction methods and real-time PCR assays

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