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Multiple Polymerase Chain Reaction for Microsporidium spp, Cryptosporidium spp, Isospora Belli and Cyclospora Cayetanensis Parasites

Received: 13 March 2014     Accepted: 17 July 2014     Published: 30 July 2014
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Abstract

The purpose of this study was to establish the optimal conditions to performance a multiple Polymerase Chain Reaction (multiplex) to detect the presence of emerging parasites Cryptosporidium spp., Mycrosporidium spp., Isospora belli, and Cyclospora spp. simultaneously, by targeting the 16S ribosomal gene in patients immunocompromised. After an exhaustive genetic analysis to obtain different primers sequences we tested the conditions of reagents concentration, time and temperature of reaction. The results showed the optimal conditions to make an efficient, specific, and reliable diagnostic method, performed in a simultaneous molecular reaction, to identify the presence of emerging parasites in faeces of patients susceptible to different infectious due to an immunocompromised status caused by diseases and chemotherapy to treat them. This method will help to decrease risk of complications and negative effects on life quality of ill patients.

Published in American Journal of Bioscience and Bioengineering (Volume 2, Issue 2)
DOI 10.11648/j.bio.20140202.12
Page(s) 33-36
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2014. Published by Science Publishing Group

Keywords

Cryptosporidium, Microsporidium, Isospora, Belli, Cyclospora, Multiplex, Immunocompromised, Emerging, Parasites

References
[1] Botero, D., Restrepo, M., Parasitosis humanas a ed. CIB, Colombia.
[2] Flanigan, T., Whalen, C., Turner, J., .Cryptosporidium infection and CD count. Ann. Intern. Med.
[3] Gonçalves, E.M., Araújo, R.S., Orban, M., Matté, G.R., Matté, M.H., Corbett, C.E. Protocol for DNA extraction of Cryptosporidium spp. oocysts in fecal samples. Rev. Inst. Med. Trop. Sao Paulo
[4] Fayer, R., Morgan, U., Upton, S.J., Epidemiology of Cryptosporidium: transmission, detection and identification. Int. J. Parasitol.
[5] Bialek, R, Binder, N., Dietz, K., Knobloch, J., Zelck, U.E. Comparison of autofluorescence and iodine staining for detection of I. belli in feces. Am. J. Trop. Med. Hyg.
[6] Mirdha, B.R., Kabra, S.K., Samantray, J.C., Isosporiasis in children. Indian. Pediatric.
[7] Orlandi, P.A., Carter, L., Brinker, A.M., da Silva, A.J., Chu, D.M., Lampel, K.A., Monday, S.R., . Targeting single-nucleotide polymorphisms in the S rRNA gene to differentiate Cyclospora species from Eimeria species by multiplex PCR. Appl. Environ. Microbiol. , -.
[8] Weitz, J.C., Weitz, C.R., Canales, M.R., Moya, R.R., Cyclospora cayetanensis infection: updated review a propos of three cases of traveler's diarrhea. Rev. Chilena. Infectol.
[9] Wasson, K., Peper, R.L., Mammalian microsporidiosis. Vet. Pathol.
[10] Viriyavejakul, P., Nintasen, R., Punsawad, C., Chaisri, U., Punpoowong, B., Riganti, M., . High prevalence of Microsporidium infection in HIV-infected patients. Southeast. Asian. J.Trop. Med. Public. Health. , -.
[11] Di Gliullo, A.B., Cribari, M.S., Bava, A.J., Cicconetti, J.S., Collazos, R., Cyclospora cayetanensis in sputum and stool samples. Rev. Inst. Med. Trop. Sao Paulo. , -.
[12] Geurden, T., Berkvens, D., Casaert, S., Vercruysse, J., Claerebout, E. A Bayesian evaluation of three diagnostic assays for the detection of Giardia duodenalis in symptomatic and asymptomatic dogs. Vet. Parasitol.
[13] Coyle, C.M., Wittner, M., Kotler, D.P., Noyer, C., Orenstein, J.M., Tanowitz, H.B., Weiss, L.M., Prevalence of microsporidiosis due to Enterocytozoon bieneusi and Encephalitozoon (Septata) intestinalis among patients with AIDS-related diarrhea: determination by polymerase chain reaction to the microsporidian small-subunit rRNA gene. Clin. Infect. Dis.
[14] Faust, E.C., D'Antoni, J.S., Odom, V., Miller, M.J., Peres, C., Sawitz, W., Thomen, L.F., Tobie, L.E., Walker, J.H., A critical study of clinical laboratory techniques for the diagnosis of protozoan cysts and helminth eggs in feces. Am. J. Trop. Med.
[15] Visvesvara, G.S., Moura. H., Kovacs-Nace, E., Wallace, S., Eberhard, M.L., Uniform staining of Cyclospora oocysts in fecal smears by a modified safranin technique with microwave heating. J. Clin. Microbiol.
[16] Dong, J., Olano, J.P., McBride, J.W., Walker, D.H., Emerging pathogens: challenges and successes of molecular diagnostics. J. Mol. Diagn.
Cite This Article
  • APA Style

    Enedina Jiménez-Cardoso, Apolinar Cano-Estrada, Adrian-Cortes Campos, Leticia Eligio-García. (2014). Multiple Polymerase Chain Reaction for Microsporidium spp, Cryptosporidium spp, Isospora Belli and Cyclospora Cayetanensis Parasites. American Journal of Bioscience and Bioengineering, 2(2), 33-36. https://doi.org/10.11648/j.bio.20140202.12

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    ACS Style

    Enedina Jiménez-Cardoso; Apolinar Cano-Estrada; Adrian-Cortes Campos; Leticia Eligio-García. Multiple Polymerase Chain Reaction for Microsporidium spp, Cryptosporidium spp, Isospora Belli and Cyclospora Cayetanensis Parasites. Am. J. BioSci. Bioeng. 2014, 2(2), 33-36. doi: 10.11648/j.bio.20140202.12

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    AMA Style

    Enedina Jiménez-Cardoso, Apolinar Cano-Estrada, Adrian-Cortes Campos, Leticia Eligio-García. Multiple Polymerase Chain Reaction for Microsporidium spp, Cryptosporidium spp, Isospora Belli and Cyclospora Cayetanensis Parasites. Am J BioSci Bioeng. 2014;2(2):33-36. doi: 10.11648/j.bio.20140202.12

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  • @article{10.11648/j.bio.20140202.12,
      author = {Enedina Jiménez-Cardoso and Apolinar Cano-Estrada and Adrian-Cortes Campos and Leticia Eligio-García},
      title = {Multiple Polymerase Chain Reaction for Microsporidium spp, Cryptosporidium spp, Isospora Belli and Cyclospora Cayetanensis Parasites},
      journal = {American Journal of Bioscience and Bioengineering},
      volume = {2},
      number = {2},
      pages = {33-36},
      doi = {10.11648/j.bio.20140202.12},
      url = {https://doi.org/10.11648/j.bio.20140202.12},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bio.20140202.12},
      abstract = {The purpose of this study was to establish the optimal conditions to performance a multiple Polymerase Chain Reaction (multiplex) to detect the presence of emerging parasites Cryptosporidium spp., Mycrosporidium spp., Isospora belli, and Cyclospora spp. simultaneously, by targeting the 16S ribosomal gene in patients immunocompromised. After an exhaustive genetic analysis to obtain different primers sequences we tested the conditions of reagents concentration, time and temperature of reaction. The results showed the optimal conditions to make an efficient, specific, and reliable diagnostic method, performed in a simultaneous molecular reaction, to identify the presence of emerging parasites in faeces of patients susceptible to different infectious due to an immunocompromised status caused by diseases and chemotherapy to treat them. This method will help to decrease risk of complications and negative effects on life quality of ill patients.},
     year = {2014}
    }
    

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    T1  - Multiple Polymerase Chain Reaction for Microsporidium spp, Cryptosporidium spp, Isospora Belli and Cyclospora Cayetanensis Parasites
    AU  - Enedina Jiménez-Cardoso
    AU  - Apolinar Cano-Estrada
    AU  - Adrian-Cortes Campos
    AU  - Leticia Eligio-García
    Y1  - 2014/07/30
    PY  - 2014
    N1  - https://doi.org/10.11648/j.bio.20140202.12
    DO  - 10.11648/j.bio.20140202.12
    T2  - American Journal of Bioscience and Bioengineering
    JF  - American Journal of Bioscience and Bioengineering
    JO  - American Journal of Bioscience and Bioengineering
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    PB  - Science Publishing Group
    SN  - 2328-5893
    UR  - https://doi.org/10.11648/j.bio.20140202.12
    AB  - The purpose of this study was to establish the optimal conditions to performance a multiple Polymerase Chain Reaction (multiplex) to detect the presence of emerging parasites Cryptosporidium spp., Mycrosporidium spp., Isospora belli, and Cyclospora spp. simultaneously, by targeting the 16S ribosomal gene in patients immunocompromised. After an exhaustive genetic analysis to obtain different primers sequences we tested the conditions of reagents concentration, time and temperature of reaction. The results showed the optimal conditions to make an efficient, specific, and reliable diagnostic method, performed in a simultaneous molecular reaction, to identify the presence of emerging parasites in faeces of patients susceptible to different infectious due to an immunocompromised status caused by diseases and chemotherapy to treat them. This method will help to decrease risk of complications and negative effects on life quality of ill patients.
    VL  - 2
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    ER  - 

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Author Information
  • Laboratory of parasitology, children hospital of Mexico, Dr. Marquez 162. Colonia Doctores, Cuauhtémoc. Mexico DF

  • Laboratory of parasitology, children hospital of Mexico, Dr. Marquez 162. Colonia Doctores, Cuauhtémoc. Mexico DF

  • Laboratory of parasitology, children hospital of Mexico, Dr. Marquez 162. Colonia Doctores, Cuauhtémoc. Mexico DF

  • Laboratory of parasitology, children hospital of Mexico, Dr. Marquez 162. Colonia Doctores, Cuauhtémoc. Mexico DF

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