Molecular detection of Trypanosoma cruzi in ocular tissue from putative corneal donors

CITTADINO E; MARÍA SUSANA LEGUIZAMÓN; GIANFRANCO ALI SANTORO; JUAN M. BURGOS; STARCENBAUM BOUCHEZ MI; FONTANA H

Mar del Plata

Congreso; XXXI Reunión Anual Sociedad Argentina de Protozoología; 2019

Infection by Trypanosoma cruzi, the etiological agent of Chagas disease, is endemic of America where 6-8 million subjects are infected. In Argentina, with about 1.5 millons of infected people T. cruzi infection prevalence is around 3.6%, index tha also affects organ donor population. Due to heart and intestines are T. cruzi target organs, seropositive individuals are exclude as donors, whereas for kidney transplant the use of both dead or living seropositive donors to negative recipients is accepted. In cornea transplants, as well as other tissues, WHO consensus makes general indications in which infected patients can receive them, whereas infected donors can be accepted, even for negative receptors, only in extreme cases and subject to the informed consent of the recipient. This general consideration is applied for safety without having been, to date, proved the parasite presence in the transplanted tissue.Herein we analyzed ocular tissues (20 corneas and sclera rings, and 7 eye muscles) from ten deceased seropositive donors (6/4 M/F, 30-74 years old) from Argentina that were admitted consecutively at Hospital Santa Lucia in Buenos Aires, Argentina. DNA extraction was carried out by means of QIAgen (DNeasy blood and tissue kit) with a previous incubation with proteinase. DNA integrity was checked by PCR amplification of the 290bp β-actin amplicon. Presence of T. cruzi DNA was analyzed by means of PCR reactions targeted to the variable region of kinetoplastid DNA (kDNA) (primers 121 and 122) and to the nuclear satellite sequence (TCZ1 and TCZ2). Considering tissue samples, 10% of corneas (2/20), 20% of sclera rings (4/20), and 14.3% of eye muscles (1/7) have positive PCR-kDNA findings. From patients analysis, corneas were T. cruzi positive in 20% (2/10) of corneas, 40% (4/10) sclera rings, and 25% (1/4) eye muscles. Interestingly, the two donors with positive corneas also had sclera positive findings, suggesting higher parasite burden or a special tissue tropism. This is the first report of T. cruzi presence in human cornea that bring light on the use of seropositive donors in transplant.