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Use of a simplified polymerase chain reaction procedure to detect Trypanosoma cruzi in blood samples from chronic chagasic patients in a rural endemic area.
- Source :
-
The American journal of tropical medicine and hygiene [Am J Trop Med Hyg] 1994 Dec; Vol. 51 (6), pp. 771-7. - Publication Year :
- 1994
-
Abstract
- 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.
- Subjects :
- Animals
Base Sequence
Brazil epidemiology
Chagas Disease blood
Chagas Disease epidemiology
Chronic Disease
DNA Primers chemistry
DNA, Kinetoplast chemistry
Humans
Molecular Sequence Data
Polymerase Chain Reaction
Rural Population
Trypanosoma cruzi genetics
Chagas Disease parasitology
DNA, Protozoan blood
Trypanosoma cruzi isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 0002-9637
- Volume :
- 51
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- The American journal of tropical medicine and hygiene
- Publication Type :
- Academic Journal
- Accession number :
- 7810810
- Full Text :
- https://doi.org/10.4269/ajtmh.1994.51.771