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Quantitative Kinetoplast DNA Assessment During Treatment of Mucosal Leishmaniasis as a Potential Biomarker of Outcome: A Pilot Study

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  • Instituto de Medicina Tropical “Alexander von Humboldt,” Universidad Peruana Cayetano Heredia, Lima, Peru; Public Health Ontario Laboratories, Public Health Ontario, Toronto, Canada; Department of Medicine, University of Toronto, Canada; Tropical Disease Unit, University Health Network Toronto General Hospital, Toronto, Canada
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Mucosal leishmaniasis (ML) is a disfiguring manifestation of Leishmania (Viannia) infection. We evaluated parasite load (PL) over time as a potential biomarker of treatment outcome in ML. PL was assessed with kinetoplast DNA quantitative real-time polymerase chain reaction (kDNA-qPCR) at enrollment, days 14 and 21–28 of therapy and 3, 6, 12–18, and 18–24 months after treatment of ML and correlated to demographic, clinical, and parasitologic factors. Forty-four patients were enrolled: 30 men and 14 women. Enrollment PL differed significantly by causative species (P < 0.001), and was higher in patients with severe ML (nasal and laryngeal involvement) compared with those with only isolated nasal involvement (median = 1,285 versus 51.5 parasites/μg tissue DNA; P = 0.005). Two patterns of PL emerged: pattern 1 (N = 23) was characterized by a sequential decline in PL during and after therapy until kDNA was undetectable. Pattern 2 (N = 18) was characterized by clearance of detectable kDNA during treatment, followed by an increased PL thereafter. All patients who failed treatment (N = 4) demonstrated pattern 1. Leishmania (Viannia) braziliensis was overrepresented among those with pattern 2 (P = 0.019). PL can be quantified by cytology brush qPCR during and after treatment in ML. We demonstrate that treatment failure was associated with undetectable PL, and L. (V.) braziliensis infection was overrepresented in those with rebounding PL.

Author Notes

* Address correspondence to Andrea K. Boggild, Tropical Disease Unit, Toronto General Hospital, 200 Elizabeth Street, 13EN-218, Toronto, ON M5G 2C4, Canada. E-mail: andrea.boggild@utoronto.ca
† These authors contributed equally to this work.

Financial support: This study was funded by the American Society of Tropical Medicine and Hygiene through the Gorgas Memorial Institute Research Award (Andrea K. Boggild). Personnel and facility support for the Arevalo molecular laboratory (Marlene Jara, Milena Alba, Vanessa Adaui, and Jorge Arevalo) was provided by the Institutional Collaboration Framework Agreement 3 from the Belgian Directorate-General for Development (project 95502). Marlene Jara was a Fogarty Scholar supported by NIH/Fogarty International Center Global Infectious Diseases Training Grant D43TW007120 during the study period.

Authors' addresses: Marlene Jara, Vanessa Adaui, Milena Alba, and Jorge Arevalo, Departmento de Bioquimica, Biologia Molecular y Farmacologia, Facultad de Ciencias, Universidad Peruana Cayetano Heredia, Lima, Peru, E-mails: jara_marlene@yahoo.es, vanessa.adaui@upch.pe, milenacollado@yahoo.es, and biomoljazz@gmail.com. Braulio Mark Valencia and Alejandro Llanos-Cuentas, Institute of Tropical Medicine Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima , Peru, E-mails: braulio.valencia@upch.pe and alejandro.llanos.c@upch.pe. Rachel Lau, Public Health Ontario Laboratories, Toronto, Canada, E-mail: rachel.lau@oahpp.ca. Andrea K. Boggild, Tropical Disease Unit, Toronto General Hospital, Toronto, Canada, E-mail: andrea.boggild@utoronto.ca.

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