Identification and detection of Trypanosoma cruzi by using a DNA amplification fingerprint obtained from the ribosomal intergenic spacer.
We designed a PCR assay targeted on repeated elements of the ribosomal intergenic spacer which produces highly polymorphic DNA band patterns for different strains of Trypanosoma cruzi. By labeling the PCR products with digoxigenin and by chemiluminescence detection, we improved the assay sensitivity by three orders of magnitude to get T. cruzi strain fingerprints in feces of the trypanosome-infected triatomine bug vector. We also developed a capture assay for the digoxigenin-labeled PCR products that allowed us to detect T. cruzi in triatomine bug vector feces and in human serum samples with a solid support.
González
N
N
Facultad de Ciencias, Universidad de Los Andes, Mérida.
Galindo
I
I
Guevara
P
P
Novak
E
E
Scorza
J V
JV
Añez
N
N
Da Silveira
J F
JF
Ramírez
J L
JL
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D015342:DNA Probes; D016054:DNA, Protozoan / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics*; D004076:Digoxigenin; D005243:Feces / Q000382:microbiology; D007303:Insect Vectors / Q000382:microbiology; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012239:Rhodnius / Q000469:parasitology; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014349:Trypanosoma cruzi / Q000235:genetics / Q000302:isolation & purification*
D015342:DNA Probes; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D004076:Digoxigenin