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Performance Evaluation of a Commercial Real-Time PCR Assay and of an In-House Real-Time PCR for Trypanosoma cruzi DNA Detection in a Tropical Medicine Reference Center, Northern Italy.


ABSTRACT: Chagas disease, a neglected protozoal disease endemic in Latin America, is also currently considered an emerging threat in nonendemic areas because of population movements. The detection of Trypanosoma cruzi DNA is increasingly being considered as important evidence to support Chagas disease diagnoses. However, further performance evaluation of molecular assays is useful for a standardization of strategy considering the whole process in routine diagnosis, especially for the different settings such as endemic and nonendemic countries. Seventy-five samples were collected from subjects screened for Chagas disease in Italy. The DNA was isolated from blood using automated extraction. We evaluated the performance of the commercial RealCycler® CHAG kit (pmPCR) based on satellite DNA (SatDNA) and of an in-house real-time PCR (ihPCR) targeting Sat and kinetoplast (k) DNAs, using the concordance of two serology assays as a reference standard. The sensitivity of kDNA and SatDNA tests by ihPCR and SatDNA by pmPCR were 14.29% (95% confidence interval (CI) 6.38 to 26.22), 7.14% (95% CI 1.98 to 17.29), and 7.14% (95% CI 1.98 to 17.29), respectively. Specificity was 100% for all PCR assays and targets. Overall, our results suggest that the preferred approach for clinical laboratories is to combine the kDNA and SatDNA as targets in order to minimize false-negative results increasing sensitivity.

SUBMITTER: Longoni SS 

PROVIDER: S-EPMC7692395 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Performance Evaluation of a Commercial Real-Time PCR Assay and of an In-House Real-Time PCR for <i>Trypanosoma cruzi</i> DNA Detection in a Tropical Medicine Reference Center, Northern Italy.

Longoni Silvia Stefania SS   Pomari Elena E   Antonelli Alberto A   Formenti Fabio F   Silva Ronaldo R   Tais Stefano S   Scarso Salvatore S   Rossolini Gian Maria GM   Angheben Andrea A   Perandin Francesca F  

Microorganisms 20201030 11


Chagas disease, a neglected protozoal disease endemic in Latin America, is also currently considered an emerging threat in nonendemic areas because of population movements. The detection of <i>Trypanosoma cruzi</i> DNA is increasingly being considered as important evidence to support Chagas disease diagnoses. However, further performance evaluation of molecular assays is useful for a standardization of strategy considering the whole process in routine diagnosis, especially for the different sett  ...[more]

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