Predictive role of polymerase chain reaction in the early diagnosis of congenital Trypanosoma cruzi infection

Velázquez, Elsa; Rivero, Rocío; Rissio, Ana María De; Malagrino, Nora; Esteva, M; Riarte, Adelina; Ruiz, Andrés M.

doi:10.1016/j.actatropica.2014.05.016

Cited by 29 publications

(27 citation statements)

References 29 publications

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“…7 In Argentina, PCR positive rates as high as 30.8% in Jujuy and 27.2% in Formosa have been recently reported. 39 PCR on cord blood is not universally accepted for diagnosing congenital Chagas disease at birth. 3 We, therefore, only used cord blood PCR negative results to identify false positive direct parasitological examinations because of reading errors.…”

Section: Discussionmentioning

confidence: 99%

Congenital Transmission of Trypanosoma cruzi in Argentina, Honduras, and Mexico: An Observational Prospective Study

Buekens

Cafferata²,

Alger³

et al. 2018

The American Journal of Tropical Medicine and Hygiene

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Compared with South America, there is a lack of epidemiologic studies about the risk of congenital transmission of in Central America and Mexico. It has been suggested that genotypes might differ by region and that congenital transmission might vary according to the parasite's genotype. Our objective was to compare congenital transmission rates in three countries. We performed an observational prospective study in 2011-2014 enrolling women at delivery in one hospital in Argentina, two hospitals in Honduras, and two hospitals in Mexico. Congenital infection was defined as the presence of one or more of the following criteria: presence of parasites in cord blood (direct parasitological microscopic examination) with positive polymerase chain reaction (PCR) in cord blood, presence of parasites in infant's blood at 4-8 weeks (direct parasitological microscopic examination), and persistence of -specific antibodies at 10 months, as measured by at least two tests. Among 28,145 enrolled women, 347 had at least one antibody rapid test positive in cord blood and a positive enzyme-linked immunosorbent assay in maternal blood. PCR in maternal blood was positive in 73.2% of the cases, and genotyping identified a majority of non-TcI in the three countries. We found no (0.0%; 95% confidence interval [CI]: 0.0, 2.0) confirmed congenital case in Honduras. Congenital transmission was 6.6% (95% CI: 3.1, 12.2) in Argentina and 6.3% (95% CI: 0.8, 20.8) in Mexico. non-TcI predominated and risks of congenital transmission were similar in Argentina and Mexico.

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Section: Discussionmentioning

confidence: 99%

Congenital Transmission of Trypanosoma cruzi in Argentina, Honduras, and Mexico: An Observational Prospective Study

Buekens

Cafferata²,

Alger³

et al. 2018

The American Journal of Tropical Medicine and Hygiene

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“…The capacity of molecular techniques, such as PCR, to detect a small amount of parasites in blood can provide an early diagnosis (2,59,60). All the current screening programs include a PCR at birth, except in Catalonia, and the test is repeated 1 month later if the result is negative, except in Valencia, where it is optional.…”

Section: Discussionmentioning

confidence: 99%

Towards a New Strategy for Diagnosis of Congenital Trypanosoma cruzi Infection

et al. 2017

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The immigration of Latin American women of childbearing age has spread the congenital transmission of Chagas disease to areas of nonendemicity, and the disease is now a worldwide problem. Some European health authorities have implemented screening programs to prevent vertical transmission, but the lack of a uniform protocol calls for the urgent establishment of a new strategy common to all laboratories. Our aims were to (i) analyze the trend of passive IgG antibodies in the newborn by means of five serological tests for the diagnosis and follow-up of congenital infection, (ii) assess the utility of these techniques for diagnosing a congenital transmission, and (iii) propose a strategy for a prompt, efficient, and cost-effective diagnosis of infection. In noninfected newborns, a continuous decreasing trend of passive IgG antibodies was observed, but none of the serological assays seroreverted in any the infants before 12 months. From 12 months onwards, serological tests achieved negative results in all the samples analyzed, with the exception of the highly sensitive chemiluminescent microparticle immunoassay (CMIA). In contrast, in congenitally infected infants, the antibody decline was detected only after treatment initiation. In order to improve the diagnosis of congenital infection, we propose a new strategy involving fewer tests that allows significant cost savings. The protocol could start 1 month after birth with a parasitological test and/or a PCR. If negative, a serological test would be carried out at 9 months, which if positive, would be followed by another at around 12 months for confirmation.

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“…Two cPCR methods were performed as described by Velázquez et al 21 : one targeting a 188-bp sequence from SatDNA and the other one targeting the 330-bp variable sequence of kDNA (Table 1). In addition, the amplification of a fragment of the ACTB gene was used to ensure the quality of DNA sample (Table 1).…”

Section: End-point Pcr Proceduresmentioning

confidence: 99%

“…In addition, the amplification of a fragment of the ACTB gene was used to ensure the quality of DNA sample (Table 1). 21 Amplifications were performed in a LifeTouch thermal cycler (BIOER, Hangzhou, China), and PCR products were visualized under a UV transilluminator after agarose gel electrophoresis using 2% agarose gels that were stained with ethidium bromide.…”

Section: End-point Pcr Proceduresmentioning

confidence: 99%

“…14,15 PCR methods seem to be more adequate for the diagnosis of congenital CD because of their high sensitivity and have been reported to be specific for the detection of T. cruzi DNA in blood samples from newborns, 16e20 demonstrating their use as a predictive tool in the diagnosis of congenital infection. 21 Several conventional end-point (cPCR) and real-time quantitative PCR (qPCR) amplification procedures to detect T. cruzi in human blood samples have been evaluated and standardized in an international multicenter study. 12 Recently, two duplex qPCR strategies based on TaqMan probes, designed to quantify both T. cruzi DNA and an internal amplification control in a single reaction tube, were subjected to an extensive analytical validation protocol and clinical evaluation with blood samples from patients with CD from different regions.…”

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Comparative Study and Analytical Verification of PCR Methods for the Diagnosis of Congenital Chagas Disease

Cura

Ramı́rez²,

Rodríguez

et al. 2017

The Journal of Molecular Diagnostics

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Congenital infection is currently the first cause of new cases of Chagas disease in Argentina and nonendemic areas worldwide. Its diagnosis is of utmost importance to guarantee curative treatment. To improve such diagnosis, a transfer process of PCR tests to the national laboratory network has been initiated. We performed a comparative study of four PCR assays [two end-point PCR and two duplex real-time quantitative PCR (qPCR) procedures] to detect Trypanosoma cruzi DNA in blood samples. Because satellite DNA and kinetoplastid DNA qPCR methods showed the best performance and the use of two different molecular targets for confirmatory purposes has been recommended, these methods were selected to perform the transfer process and, in consequence, subjected to an analytical verification protocol based on international guidelines. The anticipated reportable range was verified between 0.25 and 10 parasite equivalents per milliliter of blood (par. eq./mL) for both qPCR methods, and the limit of detection was estimated to be 0.87 (95% CI, 0.62-1.24) and 0.43 (95% CI, 0.32-0.59) par. eq./mL for satellite DNA and kinetoplastid DNA qPCR methods, respectively. In addition, both qPCR methods showed trueness and verified precision in the highest and the lowest concentrations tested. This work provides critical knowledge of the technology transfer process planned to cover laboratories of the regional network with known installed facilities.

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Predictive role of polymerase chain reaction in the early diagnosis of congenital Trypanosoma cruzi infection

Cited by 29 publications

References 29 publications

Congenital Transmission of Trypanosoma cruzi in Argentina, Honduras, and Mexico: An Observational Prospective Study

Congenital Transmission of Trypanosoma cruzi in Argentina, Honduras, and Mexico: An Observational Prospective Study

Towards a New Strategy for Diagnosis of Congenital Trypanosoma cruzi Infection

Comparative Study and Analytical Verification of PCR Methods for the Diagnosis of Congenital Chagas Disease

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