Polymerase chain reaction detection of Trypanosoma cruzi in human blood samples as a tool for diagnosis and treatment evaluation.
Trypanosoma cruzi specific sequences were amplified by the polymerase chain reaction from total blood of human chagasic patients and normal individuals. A 330 bp fragment originating from kinetoplast DNA was specifically detected in most chagasic individuals. We tested the sensitivity and specificity of this method in normal and affected individuals attending the Evandro Chagas Hospital, Rio de Janeiro. The results of these tests were compared with serological diagnosis performed using standard techniques, and in some cases with xenodiagnosis. We found that none of the serologically negative individuals gave any specific amplification product, whereas 55 out of 61 patients previously serodiagnosed as chagasic were positive using the PCR method (sensitivity: 90%). Xenodiagnosis, which is currently considered to be the most sensitive parasitological technique for Chagas' disease diagnosis, detected only 12 out of 28 serologically positive patients (sensitivity: 43%). The usefulness of the PCR method was further investigated with chagasic patients who had received anti-parasite treatment with benznidazole. It has always been difficult to evaluate the incidence of cure in such cases by serology, since a humoral response against T. cruzi antigens may remain for years even in the absence of the parasite. We observed a positive amplification result in only 9 out of 32 treated patients who remained reactive when tested using classical serology. These observations suggest that PCR is the most sensitive technique available for direct detection of T. cruzi in chagasic patients and that it can be a very useful instrument for the follow-up of patients after specific treatment.
Britto
C
C
Department of Biochemistry and Molecular Biology, Oswaldo Cruz Institute, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
Cardoso
M A
MA
Vanni
C M
CM
Hasslocher-Moreno
A
A
Xavier
S S
SS
Oelemann
W
W
Santoro
A
A
Pirmez
C
C
Morel
C M
CM
Wincker
P
P
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D001483:Base Sequence; D014355:Chagas Disease / Q000175:diagnosis* / Q000188:drug therapy / Q000276:immunology; D018105:DNA, Kinetoplast / Q000097:blood*; D006801:Humans; D008969:Molecular Sequence Data; D009593:Nitroimidazoles / Q000627:therapeutic use; D018512:Parasitemia / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014349:Trypanosoma cruzi / Q000302:isolation & purification*
D000913:Antibodies, Protozoan; D018105:DNA, Kinetoplast; D009593:Nitroimidazoles; C009999:benzonidazole