@article{fdi:010064658,
  title = {{M}ultiplex real-time {PCR} assay using {T}aq{M}an probes for the identification of {T}rypanosoma cruzi {DTU}s in biological and clinical samples},
  author = {{C}ura, {C}. {I}. and {D}uffy, {T}. and {L}ucero, {R}. {H}. and {B}isio, {M}. and {P}eneau, {J}. and {J}imenez-{C}oello, {M}. and {C}alabuig, {E}. and {G}imenez, {M}. {J}. and {A}yala, {E}. {V}. and {K}jos, {S}. {A}. and {S}antalla, {J}. and {M}ahaney, {S}. {M}. and {C}ayo, {N}. {M}. and {N}agel, {C}. and {B}arcan, {L}. and {M}achaca, {E}. {S}. {M}. and {V}iana, {K}. {Y}. {A}. and {B}rutus, {L}aurent and {O}campo, {S}. {B}. and {A}znar, {C}. and {C}uba, {C}. {A}. {C}. and {G}urtler, {R}. {E}. and {R}amsey, {J}. {M}. and {R}ibeiro, {I}. and {V}ande{B}erg, {J}. {L}. and {Y}adon, {Z}. {E}. and {O}suna, {A}. and {S}chijman, {A}. {G}.},
  editor = {},
  language = {{ENG}},
  abstract = {{B}ackground {T}rypanosoma cruzi has been classified into six {D}iscrete {T}yping {U}nits ({DTU}s), designated as {T}c{I}-{T}c{VI}. {I}n order to effectively use this standardized nomenclature, a reproducible genotyping strategy is imperative. {S}everal typing schemes have been developed with variable levels of complexity, selectivity and analytical sensitivity. {M}ost of them can be only applied cultured stocks. {I}n this context, we aimed to develop a multiplex {R}eal-{T}ime {PCR} method to identify the six {T}. cruzi {DTU}s using {T}aq{M}an probes ({MT}q-{PCR}). {M}ethods/{P}rincipal {F}indings {T}he {MT}q-{PCR} has been evaluated in 39 cultured stocks and 307 biological samples from vectors, reservoirs and patients from different geographical regions and transmission cycles in comparison with a multi-locus conventional {PCR} algorithm. {T}he {MT}q-{PCR} was inclusive for laboratory stocks and natural isolates and sensitive for direct typing of different biological samples from vectors, reservoirs and patients with acute, congenital infection or {C}hagas reactivation. {T}he first round {SL}-{IR} {MT}q-{PCR} detected 1 fg {DNA}/reaction tube of {T}c{I}, {T}c{II} and {T}c{III} and 1 pg {DNA}/reaction tube of {T}c{IV}, {T}c{V} and {T}c{VI} reference strains. {T}he {MT}q-{PCR} was able to characterize {DTU}s in 83% of triatomine and 96% of reservoir samples that had been typed by conventional {PCR} methods. {R}egarding clinical samples, 100% of those derived from acute infected patients, 62.5% from congenitally infected children and 50% from patients with clinical reactivation could be genotyped. {S}ensitivity for direct typing of blood samples from chronic {C}hagas disease patients (32.8% from asymptomatic and 22.2% from symptomatic patients) and mixed infections was lower than that of the conventional {PCR} algorithm. {C}onclusions/{S}ignificance {T}yping is resolved after a single or a second round of {R}eal-{T}ime {PCR}, depending on the {DTU}. {T}his format reduces carryover contamination and is amenable to quantification, automation and kit production.},
  keywords = {{AMERIQUE} {DU} {SUD} ; {ETATS} {UNIS}},
  booktitle = {},
  journal = {{P}los {N}eglected {T}ropical {D}iseases},
  volume = {9},
  numero = {5},
  pages = {e0003765 [18 p.]},
  ISSN = {1935-2735},
  year = {2015},
  DOI = {10.1371/journal.pntd.0003765},
  URL = {https://www.documentation.ird.fr/hor/fdi:010064658},
}