Use of a simplified polymerase chain reaction procedure to detect Trypanosoma cruzi in blood samples from chronic chagasic patients in a rural endemic area.
The feasibility of using DNA amplification by the polymerase chain reaction (PCR) for specific detection of Trypanosoma cruzi in human blood specimens was investigated. One hundred blood samples were collected in an endemic area of Minas Gerais, Brazil. They were submitted to DNA extraction and PCR amplification with kinetoplast DNA-specific primers using a simplified boiling procedure that linearized most minicircle molecules without the aid of chemical reagents. Samples that gave negative results were checked for possible inhibition of amplification using primers derived from a human-specific sequence, and those showing some level of inhibition were retested after a new DNA extraction. Of 86 patients previously diagnosed as chagasic by serologic techniques, 83 were positive in our PCR test (sensitivity = 96.5%), including all the xenodiagnosis-positive patients and 21 (87.5%) of 24 xenodiagnosis-negative individuals. In addition, four of six patients with doubtful serologic results were confirmed as positive by PCR. Our results suggest that the PCR may be a useful complement to serology in the diagnosis of Chagas' disease, and that it is the most powerful technique available for parasite detection in patients with chronic disease.
Wincker
P
P
Departamento de Bioquimica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil.
Britto
C
C
Pereira
J B
JB
Cardoso
M A
MA
Oelemann
W
W
Morel
C M
CM
D000818:Animals; D001483:Base Sequence; D001938:Brazil / Q000453:epidemiology; D014355:Chagas Disease / Q000097:blood / Q000453:epidemiology / Q000469:parasitology*; D002908:Chronic Disease; D017931:DNA Primers / Q000737:chemistry; D018105:DNA, Kinetoplast / Q000737:chemistry; D016054:DNA, Protozoan / Q000097:blood*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012424:Rural Population; D014349:Trypanosoma cruzi / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D018105:DNA, Kinetoplast; D016054:DNA, Protozoan