Limits...
Chagas disease reactivation in a heart transplant patient infected by domestic Trypanosoma cruzi discrete typing unit I (TcIDOM).

Costales JA, Kotton CN, Zurita-Leal AC, Garcia-Perez J, Llewellyn MS, Messenger LA, Bhattacharyya T, Burleigh BA - Parasit Vectors (2015)

Bottom Line: It has been previously suggested that TcI infection is benign and does not lead to chronic chagasic cardiomyopathy (CCC).Our data indicate that the parasites isolated from the patient belong to a genotype frequently associated with human infection throughout the Americas (TcIDOM).Our results constitute compelling evidence in support of TcI DTU's ability to cause end-stage CCC and help dispel any residual bias that infection with this lineage is benign, pointing to the need for increased surveillance for dissemination of this genotype in endemic regions, the USA and globally.

View Article: PubMed Central - PubMed

Affiliation: Centro de Investigación en Enfermedades Infecciosas, Escuela de Biología, Pontificia Universidad Católica del Ecuador, Avenida 12 de Octubre y Roca, Quito, Ecuador. jacostalesc@puce.edu.ec.

ABSTRACT

Background: Trypanosoma cruzi, causative agent of Chagas disease, displays high intraspecific genetic diversity: six genetic lineages or discrete typing units (DTUs) are currently recognized, termed TcI through TcVI. Each DTU presents a particular distribution pattern across the Americas, and is loosely associated with different transmission cycles and hosts. Several DTUs are known to circulate in Central America. It has been previously suggested that TcI infection is benign and does not lead to chronic chagasic cardiomyopathy (CCC).

Findings: In this study, we genotyped T. cruzi parasites circulating in the blood and from explanted cardiac tissue of an El Salvadorian patient who developed reactivation Chagas disease while on immunosuppressive medications after undergoing heart transplant in the U.S. as treatment for end-stage CCC. Parasite typing was performed through molecular methods (restriction fragment length polymorphism of polymerase reaction chain amplified products, microsatellite typing, maxicircle sequence typing and low-stringency single primer PCR, [LSSP-PCR]) as well as lineage-specific serology. We show that the parasites infecting the patient belong to the TcI DTU exclusively. Our data indicate that the parasites isolated from the patient belong to a genotype frequently associated with human infection throughout the Americas (TcIDOM).

Conclusions: Our results constitute compelling evidence in support of TcI DTU's ability to cause end-stage CCC and help dispel any residual bias that infection with this lineage is benign, pointing to the need for increased surveillance for dissemination of this genotype in endemic regions, the USA and globally.

No MeSH data available.


Related in: MedlinePlus

Molecular typing for cultured parasites and parasite clones. DNA was extracted from epimastigote hemocultures (HC) and five derived clones (CL1-CL5). TcI-TcVI correspond to DTU controls, NC corresponds PCR negative control. Lanes containing restriction products are labeled with an asterisk (*). Only restriction products are shown for controls. DNA was analyzed by the PCR-RFLP scheme proposed by Lewis, et al., 2009 [12]: as indicated by the brackets on the right side, fragments from the LSUrDNA, HSP60 and GPI genes were amplified by PCR. GPI and HSP60 products were digested with HhaI and EcoRV restriction enzimes, respectively
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4548706&req=5

Fig2: Molecular typing for cultured parasites and parasite clones. DNA was extracted from epimastigote hemocultures (HC) and five derived clones (CL1-CL5). TcI-TcVI correspond to DTU controls, NC corresponds PCR negative control. Lanes containing restriction products are labeled with an asterisk (*). Only restriction products are shown for controls. DNA was analyzed by the PCR-RFLP scheme proposed by Lewis, et al., 2009 [12]: as indicated by the brackets on the right side, fragments from the LSUrDNA, HSP60 and GPI genes were amplified by PCR. GPI and HSP60 products were digested with HhaI and EcoRV restriction enzimes, respectively

Mentions: Genotyping directly from patient’s blood samples and parafinized heart explants using a nested PCR-RFLP for the 1f8 flagellar protein and digestion with Alw21I restriction enzyme (Van der Auwera, unpublished) assigned parasites to DTU TcI (Fig. 1). Hemoculture six and eight weeks after transplant yielded epimastigotes, which were cloned in solid medium. Cultured parasites and clones were determined to belong to DTU TcI by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) as in [12] (Fig. 2). Intra-TcI genotyping was performed with nuclear microsatellites [13] and maxicircle gene fragments [14]. Intriguingly, microsatellite data indicate a close relationship with TcIDOM, a distinct genotype within DTU TcI which is common among human cases in Latin America (Fig. 3), while the maxicircle sequence analysis indicates an origin among wild/non-human isolates for North and Central America (Fig. 4). Additionally, LSSP-PCR [15] showed matching patterns among DNA extracted from patient blood samples, heart explants and cultured parasite clones, indicating that they represent the same parasite population. Meanwhile, Y-strain, from the TcII lineage, employed as a control, yielded a completely different amplification pattern (Fig. 5).Fig. 1


Chagas disease reactivation in a heart transplant patient infected by domestic Trypanosoma cruzi discrete typing unit I (TcIDOM).

Costales JA, Kotton CN, Zurita-Leal AC, Garcia-Perez J, Llewellyn MS, Messenger LA, Bhattacharyya T, Burleigh BA - Parasit Vectors (2015)

Molecular typing for cultured parasites and parasite clones. DNA was extracted from epimastigote hemocultures (HC) and five derived clones (CL1-CL5). TcI-TcVI correspond to DTU controls, NC corresponds PCR negative control. Lanes containing restriction products are labeled with an asterisk (*). Only restriction products are shown for controls. DNA was analyzed by the PCR-RFLP scheme proposed by Lewis, et al., 2009 [12]: as indicated by the brackets on the right side, fragments from the LSUrDNA, HSP60 and GPI genes were amplified by PCR. GPI and HSP60 products were digested with HhaI and EcoRV restriction enzimes, respectively
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4548706&req=5

Fig2: Molecular typing for cultured parasites and parasite clones. DNA was extracted from epimastigote hemocultures (HC) and five derived clones (CL1-CL5). TcI-TcVI correspond to DTU controls, NC corresponds PCR negative control. Lanes containing restriction products are labeled with an asterisk (*). Only restriction products are shown for controls. DNA was analyzed by the PCR-RFLP scheme proposed by Lewis, et al., 2009 [12]: as indicated by the brackets on the right side, fragments from the LSUrDNA, HSP60 and GPI genes were amplified by PCR. GPI and HSP60 products were digested with HhaI and EcoRV restriction enzimes, respectively
Mentions: Genotyping directly from patient’s blood samples and parafinized heart explants using a nested PCR-RFLP for the 1f8 flagellar protein and digestion with Alw21I restriction enzyme (Van der Auwera, unpublished) assigned parasites to DTU TcI (Fig. 1). Hemoculture six and eight weeks after transplant yielded epimastigotes, which were cloned in solid medium. Cultured parasites and clones were determined to belong to DTU TcI by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) as in [12] (Fig. 2). Intra-TcI genotyping was performed with nuclear microsatellites [13] and maxicircle gene fragments [14]. Intriguingly, microsatellite data indicate a close relationship with TcIDOM, a distinct genotype within DTU TcI which is common among human cases in Latin America (Fig. 3), while the maxicircle sequence analysis indicates an origin among wild/non-human isolates for North and Central America (Fig. 4). Additionally, LSSP-PCR [15] showed matching patterns among DNA extracted from patient blood samples, heart explants and cultured parasite clones, indicating that they represent the same parasite population. Meanwhile, Y-strain, from the TcII lineage, employed as a control, yielded a completely different amplification pattern (Fig. 5).Fig. 1

Bottom Line: It has been previously suggested that TcI infection is benign and does not lead to chronic chagasic cardiomyopathy (CCC).Our data indicate that the parasites isolated from the patient belong to a genotype frequently associated with human infection throughout the Americas (TcIDOM).Our results constitute compelling evidence in support of TcI DTU's ability to cause end-stage CCC and help dispel any residual bias that infection with this lineage is benign, pointing to the need for increased surveillance for dissemination of this genotype in endemic regions, the USA and globally.

View Article: PubMed Central - PubMed

Affiliation: Centro de Investigación en Enfermedades Infecciosas, Escuela de Biología, Pontificia Universidad Católica del Ecuador, Avenida 12 de Octubre y Roca, Quito, Ecuador. jacostalesc@puce.edu.ec.

ABSTRACT

Background: Trypanosoma cruzi, causative agent of Chagas disease, displays high intraspecific genetic diversity: six genetic lineages or discrete typing units (DTUs) are currently recognized, termed TcI through TcVI. Each DTU presents a particular distribution pattern across the Americas, and is loosely associated with different transmission cycles and hosts. Several DTUs are known to circulate in Central America. It has been previously suggested that TcI infection is benign and does not lead to chronic chagasic cardiomyopathy (CCC).

Findings: In this study, we genotyped T. cruzi parasites circulating in the blood and from explanted cardiac tissue of an El Salvadorian patient who developed reactivation Chagas disease while on immunosuppressive medications after undergoing heart transplant in the U.S. as treatment for end-stage CCC. Parasite typing was performed through molecular methods (restriction fragment length polymorphism of polymerase reaction chain amplified products, microsatellite typing, maxicircle sequence typing and low-stringency single primer PCR, [LSSP-PCR]) as well as lineage-specific serology. We show that the parasites infecting the patient belong to the TcI DTU exclusively. Our data indicate that the parasites isolated from the patient belong to a genotype frequently associated with human infection throughout the Americas (TcIDOM).

Conclusions: Our results constitute compelling evidence in support of TcI DTU's ability to cause end-stage CCC and help dispel any residual bias that infection with this lineage is benign, pointing to the need for increased surveillance for dissemination of this genotype in endemic regions, the USA and globally.

No MeSH data available.


Related in: MedlinePlus