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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: [email protected].

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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References

REFERENCES

Basuni, M., Muhi, J., Othman, N., Verweij, J. J., Ahmad, M., Miswan, N., Rahumatullah, A., Aziz, F. A., Zainudin, N. S. and Noordin, R. (2011). A pentaplex real-time polymerase chain reaction assay for detection of four species of soil transmitted helminths. American Journal of Tropical Medicine and Hygiene 84, 338343. doi: 10.4269/ajtmh.2011.10-0499.CrossRefGoogle ScholarPubMed
Concha, R., Harrington, W. Jr. and Rogers, A. I. (2005). Intestinal strongyloidiasis: recognition, management, and determinants of outcome. Journal of Clinical Gastroenterology 39, 203211.CrossRefGoogle ScholarPubMed
Dawkins, H. J. S. and Spencer, T. L. (1989). The isolation of nucleic acid from nematodes requires an understanding of the parasite and its cuticular structure. Parasitology Today 5, 7376.CrossRefGoogle ScholarPubMed
Dorris, M., Viney, M. E. and Blaxter, M. L. (2002). Molecular phylogenetic analysis of the genus Strongyloides and related nematodes. International Journal for Parasitology 32, 15071517.CrossRefGoogle ScholarPubMed
Grove, D. I. (1996). Human strongyloidiasis. Advances in Parasitology 38, 251309.CrossRefGoogle ScholarPubMed
Hasegawa, H., Hayashida, S., Ikeda, Y. and Sato, H. (2009). Hyper-variable regions in 18S rDNA of Strongyloides spp. as markers for species-specific diagnosis. Parasitology Research 104, 869874. doi: 10.1007/s00436-008-1269-9.CrossRefGoogle ScholarPubMed
Inês Ede, J., Souza, J. N., Santos, R. C., Souza, E. S., Santos, F. L., Silva, M. L., Silva, M. P., Teixeira, M. C. and Soares, N. M. (2011). Efficacy of parasitological methods for the diagnosis of Strongyloides stercoralis and hookworm in faecal specimens. Acta Tropica 3, 206210. doi: 10.1016/j.actatropica.2011.08.010.CrossRefGoogle Scholar
Janwan, P., Intapan, P. M., Thanchomnang, T., Lulitanond, V., Anamnart, W. and Maleewong, W. (2011). Rapid detection of Opisthorchis viverrini and Strongyloides stercoralis in human fecal samples using a duplex real-time PCR and melting curve analysis. Parasitology Research 109, 15931601. doi: 10.1007/s00436-011-2419-z.CrossRefGoogle ScholarPubMed
Koga, K., Kasuya, S., Khamboonruang, C., Sukhavat, K., Ieda, M., Takatsuka, N., Kita, K. and Ohtomo, H. (1991). A modified agar plate method for detection of Strongyloides stercoralis . American Journal of Tropical Medicine and Hygiene 45, 518521.CrossRefGoogle ScholarPubMed
Kramme, S., Nissen, N., Soblik, H., Erttmann, K., Tannich, E., Fleischer, B., Panning, M. and Brattig, N. J. (2011). Novel real-time PCR for the universal detection of Strongyloides species. Journal of Medical Microbiology 4, 454458. doi: 10.1099/jmm.0.025338-0.CrossRefGoogle Scholar
Kreike, J. and Lehner, A. (1995). Sex determination and DNA competition in the analysis of forensic mixed stains by PCR. International Journal of Legal Medicine 107, 235238.CrossRefGoogle ScholarPubMed
Liu, L. X. and Weller, P. F. (1993). Strongyloidiasis and other intestinal nematode infections. Infectious Disease Clinics of North America 7, 655682.CrossRefGoogle ScholarPubMed
Lutz, A. (1919). O Schistosomum mansoni e a schistosomatose segundo observações feitas no Brasil. Memórias do Instituto Oswaldo Cruz 11, 121155.CrossRefGoogle Scholar
Marcilla, A., Garg, G., Bernal, D., Ranganathan, S., Forment, J., Ortiz, J., Muñoz-Antolí, C., Dominguez, M. V., Pedrola, L., Martinez-Blanch, J., Sotillo, J., Trelis, M., Toledo, R. and Esteban, J. G. (2012). The transcriptome analysis of Strongyloides stercoralis L3i larvae reveals targets for intervention in a neglected disease. PLoS Neglected Tropical Diseases 6, e1513. doi: 10.1371/journal.pntd.0001513.CrossRefGoogle Scholar
Marcos, L. A., Terashima, A., DuPont, H. L. and Gotuzzo, E. (2008). Strongyloides hyperinfection syndrome: an emerging global infectious disease. Transactions of the Royal Society of Tropical Medicine and Hygiene 102, 314318. doi: 10.1007/s11908-010-0150-z.CrossRefGoogle ScholarPubMed
Marra, N. M., Chiuso-Minicucci, F., Machado, G. C., Zorzella-Pezavento, S. F., França, T. G., Ishikawa, L. L., Amarante, A. F., Sartori, A. and Amarante, M. R. (2010). Faecal examination and PCR to detect Strongyloides venezuelensis in experimentally infected Lewis rats. Memórias do Instituto Oswaldo Cruz 105, 5761. doi: 10.1590/S0074-02762010000100008.CrossRefGoogle ScholarPubMed
Mejia, R. and Nutman, T. B. (2012). Screening, prevention, and treatment for hyperinfection syndrome and disseminated infections caused by Strongyloides stercoralis . Current Opinion in Infectious Diseases 4, 458463. doi: 10.1097/QCO.0b013e3283551dbd.CrossRefGoogle Scholar
Moghaddassani, H., Mirhendi, H., Hosseini, M., Rokni, M., Mowlavi, G. and Kia, E. (2011). Molecular diagnosis of Strongyloides stercoralis infection by PCR detection of specific DNA in human stool samples. Iranian Journal of Parasitology 6, 2330.Google ScholarPubMed
Nilforoushan, M. R., Mirhendi, H., Rezaie, S., Rezaian, M., Meamar, A. R. and Kia, E. B. (2007). A DNA-based identification of Strongyloides stercoralis isolates from Iran. Iranian Journal of Public Health 36, 620.Google Scholar
Olsen, A., van Lieshout, L., Marti, H., Polderman, T., Polman, K., Steinmann, P., Stothard, R., Thybo, S., Verweij, J. J. and Magnussen, P. (2009). Strongyloidiasis – the most neglected of the neglected tropical diseases? Transactions of the Royal Society of Tropical Medicine and Hygiene 103, 967972. doi: 10.1016/j.trstmh.2009.02.013.CrossRefGoogle ScholarPubMed
Paula, F. M. and Costa-Cruz, J. M. (2011). Epidemiological aspects of strongyloidiasis in Brazil. Parasitology 138, 13311340. doi: 10.1017/S003118201100120X.CrossRefGoogle ScholarPubMed
Repetto, S. A., Soto, C. D., Cazorla, S. I., Tayeldin, M. L., Cuello, S., Lasala, M. B., Tekiel, V. S. and González Cappa, S. M. (2013). An improved DNA isolation technique for PCR detection of Strongyloides stercoralis in stool samples. Acta Tropica 126, 110114. doi: 10.1016/j.actatropica.2013.02.003.CrossRefGoogle ScholarPubMed
Rugai, E., Mattos, T. and Brisola, A. P. (1954). Nova técnica para isolar larvas de nematóides das fezes: modificação do método de Baermann. Revista do Instituto Adolfo Lutz 14, 58.Google ScholarPubMed
Schär, F., Odermatt, P., Khieu, V., Panning, M., Duong, S., Muth, S., Marti, H. and Kramme, S. (2013). Evaluation of real-time PCR for Strongyloides stercoralis and hookworm as diagnostic tool in asymptomatic schoolchildren in Cambodia. Acta Tropica 126, 8992. doi: 10.1016/j.actatropica.2012.12.012.CrossRefGoogle ScholarPubMed
Siddiqui, A. A. and Berk, S. L. (2001). Diagnosis of Strongyloides stercoralis infection. Clinical Infectious Diseases 7, 10401047.CrossRefGoogle Scholar
Stensvold, C. R., Lebbad, M. and Verweij, J. J. (2011). The impact of genetic diversity in protozoa on molecular diagnostics. Trends in Parasitology 27, 5358. doi: 10.1016/j.pt.2010.11.005.CrossRefGoogle ScholarPubMed
Taniuchi, M., Verweij, J. J., Noor, Z., Sobuz, S. U., Lieshout, L., Petri, W. A. Jr., Haque, R. and Houpt, E. R. (2011). High through put multiplex PCR and probe-based detection with Luminex beads for seven intestinal parasites. American Journal of Tropical Medicine and Hygiene 84, 332337. doi: 10.4269/ajtmh.2011.10-0461.CrossRefGoogle Scholar
ten Hove, R. J., van Esbroeck, M., Vervoort, T., van den Ende, J., van Lieshout, L. and Verweij, J. J. (2009). Molecular diagnostics of intestinal parasites in returning travellers. European Journal of Clinical Microbiology and Infectious Diseases 28, 10451053. doi: 10.1007/s10096-009-0745-1.CrossRefGoogle ScholarPubMed
Uparanukraw, P., Phongsri, S. and Morakote, N. (1999). Fluctuations of larval excretion in Strongyloides stercoralis infection. American Journal of Tropical Medicine and Hygiene 60, 967973.CrossRefGoogle ScholarPubMed
Verweij, J. J., Canales, M., Polman, K., Ziem, J., Brienen, E. A., Polderman, A. M. and van Lieshout, L. (2009). Molecular diagnosis of Strongyloides stercoralis in faecal samples using real-time PCR. Transactions of the Royal Society of Tropical Medicine and Hygiene 103, 342346. doi: 10.1016/j.trstmh.2008.12.001.CrossRefGoogle ScholarPubMed