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Conventional PCR for molecular diagnosis of human strongyloidiasis

Published online by Cambridge University Press:  28 January 2014

R. B. SITTA
Affiliation:
Instituto de Medicina Tropical, Universidade de São Paulo, São Paulo, Brazil
F. M. MALTA
Affiliation:
Instituto de Medicina Tropical, Universidade de São Paulo, São Paulo, Brazil Departamento de Gastroenterologia (LIM/07), Faculdade de Medicina, Hospital das Clínicas, Universidade de São Paulo, São Paulo, Brazil
J. R. PINHO
Affiliation:
Instituto de Medicina Tropical, Universidade de São Paulo, São Paulo, Brazil Departamento de Gastroenterologia (LIM/07), Faculdade de Medicina, Hospital das Clínicas, Universidade de São Paulo, São Paulo, Brazil
P. P. CHIEFFI
Affiliation:
Instituto de Medicina Tropical, Universidade de São Paulo, São Paulo, Brazil Faculdade de Ciências Médicas, Santa Casa, São Paulo, Brazil
R. C. B. GRYSCHEK
Affiliation:
Departamento de Moléstias Infecciosas e Parasitárias (LIM/06), Faculdade de Medicina, Hospital das Clínicas, Universidade de São Paulo, São Paulo, Brazil
F. M. PAULA*
Affiliation:
Departamento de Moléstias Infecciosas e Parasitárias (LIM/06), Faculdade de Medicina, Hospital das Clínicas, Universidade de São Paulo, São Paulo, Brazil
*
*Corresponding author. Laboratório de Investigação Médica (LIM/06), Instituto de Medicina Tropical, Universidade de São Paulo, Av. Dr. Enéas de Carvalho Aguiar, 470, 05403-000 São Paulo, SP, Brazil. E-mail: fabiana.paula@hc.fm.usp.br.

Summary

Strongyloidiasis is frequently asymptomatic and diagnosis of latent infection is difficult due to limitations of current parasitological and serological methods. This study aimed to verify the use of conventional polymerase chain reaction (PCR) assay for molecular diagnosis of Strongyloides stercoralis infection. Fresh stool samples were obtained from 103 individuals: 33 S. stercoralis positive, 30 positive for other parasites and 40 negative for parasitological methods. These samples were examined by the Lutz, Rugai and agar plate culture methods and conventional PCR assay. Two sets of primers (S. stercoralis species-specific and genus-specific sets), located in the 18S ribosomal RNA gene, were used for PCR. Of the 33 samples positive for S. stercoralis by parasitological methods, 28 (84·8%) were also detected by PCR assay using species-specific primers and 26 (78·8%) using genus-specific primers. Among the stool samples negative by parasitological methods, seven (17·5%) were positive by PCR using species-specific primers and two (5·0%) using genus-specific primers. In conclusion, the conventional PCR assay described in this study using a species-specific primer pair provided a molecular method for S. stercoralis diagnosis in human stool samples.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2014 

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