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Early polymerase chain reaction detection of Chagas disease reactivation in heart transplant patients  Priscilla Almeida da Costa, MS, Marcela Segatto,

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Presentation on theme: "Early polymerase chain reaction detection of Chagas disease reactivation in heart transplant patients  Priscilla Almeida da Costa, MS, Marcela Segatto,"— Presentation transcript:

1 Early polymerase chain reaction detection of Chagas disease reactivation in heart transplant patients  Priscilla Almeida da Costa, MS, Marcela Segatto, PhD, Danielle Fernandes Durso, PhD, Wagson José de Carvalho Moreira, BSc, Lucas Lodi Junqueira, MD, PhD, Fábio Morato de Castilho, MD, MS, Silvio Amadeu de Andrade, MD, Cláudio Léo Gelape, MD, PhD, Egler Chiari, PhD, Andréa Teixeira-Carvalho, PhD, Sergio Danilo Junho Pena, MD, PhD, Carlos Renato Machado, PhD, Gloria Regina Franco, PhD, Geraldo Brasileiro Filho, MD, PhD, Maria da Consolação Vieira Moreira, MD, PhD, Andréa Mara Macedo, PhD  The Journal of Heart and Lung Transplantation  Volume 36, Issue 7, Pages (July 2017) DOI: /j.healun Copyright © 2017 International Society for the Heart and Lung Transplantation Terms and Conditions

2 Figure 1 Representative results of the PCR techniques. (A) Amplification of variable 330/660-bp regions of the T cruzi kDNA minicircle from endomyocardial biopsies of heart transplant patients: 1 to 12: EMB specimens of patients, where 4 showed positive amplification products; 13: DNA from T cruzi JG strain (positive control); 14: no DNA template (negative control). MW, standard molecular weight (1-kb Plus DNA Ladder; Invitrogen). (B) Dissociation curves obtained by the melting curve program for the rDNA 24Sα gene from biopsies of heart transplant patients. (I) Unspecific peaks obtained from negative EMB specimens in contrast to negative controls (without DNA). (II) Typical peaks derived from T cruzi DNA of JG (TcII) and Col1.7G2 (TcI) populations (positive controls). (III) Peaks obtained from positive EMB specimens in contrast to positive controls. The Journal of Heart and Lung Transplantation  , DOI: ( /j.healun ) Copyright © 2017 International Society for the Heart and Lung Transplantation Terms and Conditions

3 Figure 2 Comparison of reactivation results between clinical and PCR molecular diagnostic procedures using EMB specimens (paraffin-embedded or fresh biopsies) from heart transplant patients with chagasic cardiopathy. (1) aTrue positive; bfalse positive; cfalse negative; dtrue negative; (2) N = 58; (3) total number of positive tests (a + b) = 39; (4) total number of clinical reactivations (a + c) = 38. The Journal of Heart and Lung Transplantation  , DOI: ( /j.healun ) Copyright © 2017 International Society for the Heart and Lung Transplantation Terms and Conditions

4 Figure 3 ROC curves for different diagnosis approaches compared with clinical reactivation of Chagas disease. (A) Molecular diagnosis and histopathologic analysis of 500 EMB specimens from 58 patients (B) Molecular diagnosis, histopathologic analysis, blood smear and hemoculture of samples from 29 patients. Criterion >0 (positive = 1, negative = 0); AUC, area under the receiver operating characteristic (ROC) curve (p ≤ 0.001). The Journal of Heart and Lung Transplantation  , DOI: ( /j.healun ) Copyright © 2017 International Society for the Heart and Lung Transplantation Terms and Conditions

5 Figure 4 Proposed algorithm for managing chagasic patients submitted to EMB. Number of patients associated to each box represents the number of cases observed in the present study. CDR, Chagas disease reactivation; EMB, endomyocardial biopsy. The Journal of Heart and Lung Transplantation  , DOI: ( /j.healun ) Copyright © 2017 International Society for the Heart and Lung Transplantation Terms and Conditions

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