Evaluation of Molecular Methods to Identify Chagas Disease and Leishmaniasis in Blood Donation Candidates in Two Brazilian Centers

• Published in: Pathogens (Basel) 2023-03, Vol.12 (4), p.508
• Main Authors: Ferreira, Juliana de Jesus Guimarães, Costa, Sandra Cecília Botelho, Addas-Carvalho, Marcelo, Pereira, Mariane Barroso, França, Adriana de Oliveira, de Lima, Rodrigo Gonçalves, Andrade, Paula Durante, Wanderley, Jamiro da Silva, Martins, Luiz Cláudio, de Almeida, Eros Antonio, Marcon, Gláucia Elisete Barbosa
• Format: Article
• Language: English
• Subjects: Analysis; Antibodies; Antigens; Blood; blood donation; Blood donors; Care and treatment; Chagas disease; Chemiluminescence; Diagnosis; Enzyme-linked immunosorbent assay; Epidemiology; Fluorescence; Fluorescent antibody test; Health aspects; Identification methods; Immunoassay; Indirect fluorescent antibody test; Leishmania; Leishmaniasis; Melt temperature; Melting; melting curve analysis; Microparticles; Molecular dynamics; nPCR; Parasites; Parasitic diseases; PCR; Polymerase chain reaction; Protozoa; Screening; Serological tests; Serology; Trypanosoma cruzi; Vector-borne diseases
• ISSN: 2076-0817; 2076-0817
• DOI: 10.3390/pathogens12040508

In Brazil, blood donation is regulated by the Brazilian Ministry of Health, and all States follow the same protocol for clinical and laboratory screening. Brazil is an endemic country for Chagas disease (CD), caused by cruzi, and for leishmaniasis, caused by a species of spp. Screening for leishmaniosis is not routinely performed by blood banks. Given the antigenic similarity between and spp., cross-reactions in serological tests can occur, and inconclusive results for CD have been found. The objective of this study was to apply molecular techniques, e.g., nPCR, PCR, and qPCR, to clarify cases of blood donation candidates with non-negative serology for CD and to analyze the difference between the melting temperature during real-time PCR using SYBR Green. Thirty-seven cases that showed non-negative results for CD using chemiluminescent microparticle immunoassay (CMIA) tests from blood banks in Campo Grande, MS, and Campinas, SP, were analyzed. In the serum samples, 35 samples were evaluated by ELISA, and 24.3% (9/35) showed positive results for CD. nPCR was able to detect 12 positive results in 35 samples (34.28%). qPCR for was quantifiable in the samples that showed a value ≥0.002 par eq/mL (parasite equivalents per milliliter), and in 35 samples, 11 (31.42%) were positive. Of all evaluated samples using the described tests (CMIA, ELISA, nPCR, and qPCR), 18 (48.6%) were positive for CD. For MCA by qPCR, the melting temperature was 82.06 °C ± 0.46 for and 81.9 °C ± 0.24 for . The Mann-Whitney test showed a significant value of < 0.0001. However, the differentiation between and could not be considered due to temperature overlap. For leishmaniasis, of the 35 samples with non-negative serology for CD tested by the indirect fluorescent antibody test (IFAT), only one sample (2.85%) was positive (1:80). The PCR for spp. was performed on 36 blood samples from donation candidates, and all were negative. qPCR for showed 37 negative results for the 37 analyzed samples. The data presented here show the importance of performing two different tests in CD screening at blood banks. Molecular tests should be used for confirmation, thereby improving the blood donation system.