Molecular Diagnosis of Chagas Disease in Patients with Megaesophagus Exhibiting Negative or Inconclusive Serological Results

Angelica Martins Batista¹Tycha Bianca Sabaini Pavan^2*Eros Antonio de Almeida³Daniel Maximo Correa Alcantara⁴Paula Durante Andrade⁵Luiz Claudio Martins³Jamiro da Silva Wanderley³Sandra Cecila Botelho Costa⁶Glaucia Elisete Barbosa Marcon^4,5*

• ¹Center of Biological and Health Sciences, Estácio de Sá University, Rio de Janeiro RJ, Brazil., Rio de Janeiro, Brazil
• ²Gonçalo Moniz Institute (IGM), Salvador, Brazil
• ³Grupo de Estudos em Doença de Chagas (GEDoCh), Department of Medical Clinic, Faculty of Medical Sciences, State University of Campinas, Campinas, São Paulo SP, Brazil, Campinas, Brazil
• ⁴Oswaldo Cruz Foundation Mato Grosso do Sul, Campo Grande MS, Brazil, Mato Grosso do Sul, Brazil
• ⁵Laboratory of Diagnosis of Infectious Diseases by Molecular Biology Techniques, Faculty of Medical Sciences, State University of Campinas SP, Brazil, Campinas, Brazil
• ⁶Laboratory of Diagnosis of Infectious Diseases by Molecular Biology Techniques, Faculty of Medical Sciences, State University of Campinas SP, Brazil., Campinas, Brazil

Chagas disease (CD), and it has not been extensively documented in literature. However, individuals may exhibit negative or inconclusive serology for CD. This study aimed to assess the performance of molecular diagnostics for CD in participants with these conditions. This was a prospective cohort study that included 26 participants with negative or inconclusive conventional CD serology (Group I), 33 participants with positive CD serology and megaesophagus (Group II), and 10 participants with negative serology and no CD epidemiological history (Group III). Blood samples were collected for serological tests (ELISA and IFAT), blood cultures, and molecular tests like nested PCR (nPCR) targeting Sat-DNA and kDNA, as well as quantitative PCR (qPCR) of T. cruzi. Statistical analyses applying the Composite Reference Standard (CRS), showed that diagnosis by nPCR satDNA had a sensitivity of 95% (95% CI: 82%-99%), a specificity of 81% (95% CI: 64%-93%), an accuracy of 88%. When considering a positive result from at least one molecular test, 20 out of 26 participants with megaesophagus and negative or inconclusive conventional serology were identified (76.9%).This study reinforce the greater detection capacity of Sat-DNA nPCR compared to the diagnostic methods tested. This emphasizes the importance of employing molecular diagnosis to clarify the etiology in megaesophagus cases.