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Reactivation of Chagas Disease in a Patient With Follicular Lymphoma Diagnosed by Means of Quantitative Real-Time Polymerase Chain Reaction.

Garzón MI, Sánchez AG, Goy MC, Alvarellos T, Zarate AH, Basquiera AL, Garcia JJ, Caeiro JP - Open Forum Infect Dis (2015)

Bottom Line: Parasitic load was 577 950 parasite equivalents/mL.The patient began treatment with benznidazole 5 mg/k per day every 12 hours.The patient completed 60 days of treatment and is currently asymptomatic.

View Article: PubMed Central - PubMed

Affiliation: Infectious Diseases Section.

ABSTRACT
We report a case of Chagas disease reactivation in a patient with stage IIb follicular lymphoma in the cecum. He was admitted to the hospital with neutropenia and fever. He had a history of right hemicolectomy 6 months earlier and had received the sixth cycle of chemotherapy with cyclophosphamide/doxorubicin/vincristine/prednisone/rituximab. Blood and urine cultures were negative, but the fever persisted. Reactivation of Chagas disease was confirmed by means of quantitative real-time polymerase chain reaction (qRT-PCR). Parasitic load was 577 950 parasite equivalents/mL. The patient began treatment with benznidazole 5 mg/k per day every 12 hours. After 1 month, the qRT-PCR control was undetectable. The patient completed 60 days of treatment and is currently asymptomatic. Trypanosoma cruzi qRT-PCR may become a useful diagnostic method for reactivation of Chagas disease.

No MeSH data available.


Related in: MedlinePlus

(A) Amplification curve of Trypanosoma cruzi before treatment. Cycle threshold: 23 for patient and 29 for standard. (B) Amplification curve of T cruzi after treatment. Cycle threshold: 28.9 for standard. Abbreviations: N, no template control (NTC); P, patient, duplicate samples; S, standard (4500 parasite equivalents/mL).
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OFV060F1: (A) Amplification curve of Trypanosoma cruzi before treatment. Cycle threshold: 23 for patient and 29 for standard. (B) Amplification curve of T cruzi after treatment. Cycle threshold: 28.9 for standard. Abbreviations: N, no template control (NTC); P, patient, duplicate samples; S, standard (4500 parasite equivalents/mL).

Mentions: A 60 year-old male with history of stage IIb follicular lymphoma in the cecum was admitted to our institution for febrile neutropenia and asthenia. He underwent right hemicolectomy 6 months earlier and received the sixth cycle of chemotherapy with cyclophosphamide/doxorubicin/vincristine/prednisone/rituximab 1 week prior. Upon admission, he received empirical treatment with piperacillin-tazobactam after blood cultures were drawn, but the fever persisted. A thoracic computed tomography scan was performed without significant findings. The patient had history of positive Chagas serology 5 years earlier. On the eighth day of hospitalization, he remained with fever and asthenia but without positive cultures or response to antibiotics. New blood cultures for bacteria and fungi were obtained. A blood smear searching for parasites and Chagas quantitative real-time PCR (qRT-PCR) was carried out. Parasites were not observed in Giemsa-stained blood smear. The qRT-PCR for Chagas was 577 950 parasite equivalents (Par Eq)/mL (Figure 1A). Nucleic acids were isolated from 400 µL peripheral blood through MagNA Pure Compact Nucleic Acid Isolation kit 1 (Roche) and eluted in 100 µL. Quantitative real-time PCR was performed according to Piron [5] with modifications, using 2X Universal KAPA Master mix (Kapabiosystems), 10 µM TaqMan probe (Applied Biosystems), 750 nM of each primer, and 5 µL DNA sample, on a real-time thermocycler (Rotor-gene 6000; Corbett Research), with the aim of amplifying a nuclear satellite fragment of 166 base pairs from repeated region of parasitic satellite DNA. To generate a standard curve, DNA was extracted from blood sample spiked with Y strain trypomastigotes culture (Ref. number Medline: 97179491) as described previously [6]. Parasite load was determined using an absolute quantification method with a linear range of 50–1 000 000 Par Eq/mL. To validate this procedure, 1 standard of quantification (4500 Par Eq/mL), which represents a point in the standard curve accomplished earlier, and nontemplate control were included in the run.Figure 1.


Reactivation of Chagas Disease in a Patient With Follicular Lymphoma Diagnosed by Means of Quantitative Real-Time Polymerase Chain Reaction.

Garzón MI, Sánchez AG, Goy MC, Alvarellos T, Zarate AH, Basquiera AL, Garcia JJ, Caeiro JP - Open Forum Infect Dis (2015)

(A) Amplification curve of Trypanosoma cruzi before treatment. Cycle threshold: 23 for patient and 29 for standard. (B) Amplification curve of T cruzi after treatment. Cycle threshold: 28.9 for standard. Abbreviations: N, no template control (NTC); P, patient, duplicate samples; S, standard (4500 parasite equivalents/mL).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4498252&req=5

OFV060F1: (A) Amplification curve of Trypanosoma cruzi before treatment. Cycle threshold: 23 for patient and 29 for standard. (B) Amplification curve of T cruzi after treatment. Cycle threshold: 28.9 for standard. Abbreviations: N, no template control (NTC); P, patient, duplicate samples; S, standard (4500 parasite equivalents/mL).
Mentions: A 60 year-old male with history of stage IIb follicular lymphoma in the cecum was admitted to our institution for febrile neutropenia and asthenia. He underwent right hemicolectomy 6 months earlier and received the sixth cycle of chemotherapy with cyclophosphamide/doxorubicin/vincristine/prednisone/rituximab 1 week prior. Upon admission, he received empirical treatment with piperacillin-tazobactam after blood cultures were drawn, but the fever persisted. A thoracic computed tomography scan was performed without significant findings. The patient had history of positive Chagas serology 5 years earlier. On the eighth day of hospitalization, he remained with fever and asthenia but without positive cultures or response to antibiotics. New blood cultures for bacteria and fungi were obtained. A blood smear searching for parasites and Chagas quantitative real-time PCR (qRT-PCR) was carried out. Parasites were not observed in Giemsa-stained blood smear. The qRT-PCR for Chagas was 577 950 parasite equivalents (Par Eq)/mL (Figure 1A). Nucleic acids were isolated from 400 µL peripheral blood through MagNA Pure Compact Nucleic Acid Isolation kit 1 (Roche) and eluted in 100 µL. Quantitative real-time PCR was performed according to Piron [5] with modifications, using 2X Universal KAPA Master mix (Kapabiosystems), 10 µM TaqMan probe (Applied Biosystems), 750 nM of each primer, and 5 µL DNA sample, on a real-time thermocycler (Rotor-gene 6000; Corbett Research), with the aim of amplifying a nuclear satellite fragment of 166 base pairs from repeated region of parasitic satellite DNA. To generate a standard curve, DNA was extracted from blood sample spiked with Y strain trypomastigotes culture (Ref. number Medline: 97179491) as described previously [6]. Parasite load was determined using an absolute quantification method with a linear range of 50–1 000 000 Par Eq/mL. To validate this procedure, 1 standard of quantification (4500 Par Eq/mL), which represents a point in the standard curve accomplished earlier, and nontemplate control were included in the run.Figure 1.

Bottom Line: Parasitic load was 577 950 parasite equivalents/mL.The patient began treatment with benznidazole 5 mg/k per day every 12 hours.The patient completed 60 days of treatment and is currently asymptomatic.

View Article: PubMed Central - PubMed

Affiliation: Infectious Diseases Section.

ABSTRACT
We report a case of Chagas disease reactivation in a patient with stage IIb follicular lymphoma in the cecum. He was admitted to the hospital with neutropenia and fever. He had a history of right hemicolectomy 6 months earlier and had received the sixth cycle of chemotherapy with cyclophosphamide/doxorubicin/vincristine/prednisone/rituximab. Blood and urine cultures were negative, but the fever persisted. Reactivation of Chagas disease was confirmed by means of quantitative real-time polymerase chain reaction (qRT-PCR). Parasitic load was 577 950 parasite equivalents/mL. The patient began treatment with benznidazole 5 mg/k per day every 12 hours. After 1 month, the qRT-PCR control was undetectable. The patient completed 60 days of treatment and is currently asymptomatic. Trypanosoma cruzi qRT-PCR may become a useful diagnostic method for reactivation of Chagas disease.

No MeSH data available.


Related in: MedlinePlus