Resolution of multiclonal infections of Trypanosoma cruzi from naturally infected triatomine bugs and from experimentally infected mice by direct plating on a sensitive solid medium.
The isolation of biological clones of Trypanosoma cruzi by microscopically dispensing individual organisms or by serial dilution is laborious and time consuming. The inability to resolve mixed T. cruzi infections, from vectors and hosts, and to isolate clones of slow growing genotypes by efficient plating on solid media, has hindered characterisation studies and downstream applications. We have devised and validated a sensitive, solid medium plating technique for rapid in vitro isolation of clones representative of all the recognised T. cruzi lineages (TCI, TCIIa-e), including the slow growing strain CANIII (TC IIa) and Trypanosoma rangeli, with high plating efficiencies. Furthermore, the method is effective for the isolation of clones directly from silvatic triatomine bugs and from experimentally infected mice harbouring mixed infections, allowing resolution of multiclonal infections from varied sources.
Yeo
Matthew
M
Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London, WC1E 7HT, UK. Matthew.yeo@lshtm.ac.uk
Lewis
Michael D
MD
Carrasco
Hernan J
HJ
Acosta
Nidia
N
Llewellyn
Martin
M
da Silva Valente
Sebastião Aldo
SA
de Costa Valente
Vera
V
de Arias
Antonieta Rojas
AR
Miles
Michael A
MA
D000818:Animals; D014355:Chagas Disease / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D003470:Culture Media; D016054:DNA, Protozoan / Q000032:analysis; D007303:Insect Vectors / Q000469:parasitology; D051379:Mice; D018512:Parasitemia / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods; D012239:Rhodnius / Q000469:parasitology; D014224:Triatoma / Q000469:parasitology; D014225:Triatominae / Q000469:parasitology*; D014349:Trypanosoma cruzi / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification
D003470:Culture Media; D016054:DNA, Protozoan