title: Use of polymerase chain reaction (PCR) and hybridization assays to detect Trypanosoma cruzi in chronic chagasic patients treated with itraconazole or allopurinol.
abstract: The presence of Trypanosoma cruzi in chronic chagasic patients with negative xenodiagnosis (XD) after 6 years following completion of therapy with either itraconazole or allopurinol was assessed by polymerase chain reaction (PCR) and hybridization assays. A 330-bp DNA fragment amplified from the hypervariable regions of T. cruzi kinetoplastid minicircles was hybridized with total 32P-labeled kinetoplast DNA as probes. PCR alone enabled the identification of T. cruzi nucleotide sequences in 40% of the patients treated with itraconazole and in 60% of patients treated with allopurinol. PCR used in combination with hybridization detected parasite DNA in 60% and 53% of XD negative individuals treated with itraconazole or allopurinol, respectively. These results show that PCR and hybridization are more sensitive than conventional parasitological techniques in diagnosing patients that have undergone chemotherapy with itraconazole or allopurinol.
authors.affiliation: Cellular and Molecular Biology Program, Biomedical Sciences Institute, Faculty of Medicine, University of Chile, Santiago, Chile. izulanta@machi.med.uchile.cl
authors.forename: Inés
authors.initials: I
authors.lastname: Zulantay
authors.forename: Patricia
authors.initials: P
authors.lastname: Honores
authors.forename: Aldo
authors.initials: A
authors.lastname: Solari
authors.forename: Werner
authors.initials: W
authors.lastname: Apt
authors.forename: Sylvia
authors.initials: S
authors.lastname: Ortiz
authors.forename: Antonio
authors.initials: A
authors.lastname: Osuna
authors.forename: Antonio
authors.initials: A
authors.lastname: Rojas
authors.forename: Bernardo
authors.initials: B
authors.lastname: López
authors.forename: Gittith
authors.initials: G
authors.lastname: Sánchez
mesh_terms: D000328:Adult; D000493:Allopurinol / Q000494:pharmacology* / Q000627:therapeutic use; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology* / Q000627:therapeutic use; D014355:Chagas Disease / Q000097:blood / Q000175:diagnosis / Q000188:drug therapy / Q000469:parasitology*; D002908:Chronic Disease; D005260:Female; D006801:Humans; D017964:Itraconazole / Q000494:pharmacology* / Q000627:therapeutic use; D008297:Male; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D000981:Antiprotozoal Agents; D017964:Itraconazole; D000493:Allopurinol