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Comparative analysis of routine parasitological methods for recovery of cysts, molecular detection, and genotyping of Giardia duodenalis

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Abstract

In order to improve the diagnosis of giardiasis, fecal samples (high/medium/low concentration of cysts) were processed by the parasitological methods used in the routine: Faust, Lutz e Ritchie modified (replacement of formaldehyde by distilled water). The cysts were quantified; the DNA was extracted and amplified by semi-nested PCR (GDH gene). Fifteen clinical samples were analyzed to validate the study by PCR-RFLP. The results showed that the parasite was only detected and genotyped correctly when samples from children with high, medium, and low parasitic load, belonging to genotype AII, were processed by the modified Ritchie method, different from what was observed for the other methods used in laboratory routine (Faust and Lutz). The modified Ritchie method proved to be more suitable, recovering a greater number of cysts from samples, regardless of parasitic load, which reduces the chance of false negative results and has epidemiological repercussions since individuals with low parasite load are usually asymptomatic and the main disseminators of this infection.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001.

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Authors and Affiliations

  1. Postgraduate Program in Health Sciences, State University of Maringá, Maringá, Paraná, Brazil

    Renata Coltro Bezagio, Liara Izabela Lopes Romera, Caroline Rodrigues de Almeida & Mônica Lúcia Gomes

  2. Department of Basic Health Sciences, State University of Maringá, Maringá, Paraná, Brazil

    Cristiane Maria Colli & Mônica Lúcia Gomes

  3. Department of Statistics, State University of Maringá, Maringá, Paraná, Brazil

    Érika Cristina Ferreira

Authors
  1. Renata Coltro Bezagio
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  2. Cristiane Maria Colli
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  3. Liara Izabela Lopes Romera
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  4. Caroline Rodrigues de Almeida
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  5. Érika Cristina Ferreira
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  6. Mônica Lúcia Gomes
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Contributions

The experiments were designed by Renata Coltro Bezagio, Cristiane Maria Colli and Mônica Lúcia Gomes. Data collection, material preparation, and experiments were performed by Renata Coltro Bezagio, Cristiane Maria Colli, Caroline Rodrigues de Almeida, and Liara Izabela Lopes Romera. Data analysis was performed by Renata Coltro Bezagio, Érika Cristina Ferreira, and Mônica Lúcia Gomes. The first draft of the manuscript was written by Renata Coltro Bezagio, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript

Corresponding author

Correspondence to Renata Coltro Bezagio.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki Declaration. The stool samples used in this study came from a project previously approved by the Human Research Ethics Committee of the Integrado University Center of Campo Mourão-Paraná/Brazil (registration n° 1.594.078).

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Informed consent was obtained from all those responsible for the children included in this study.

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Bezagio, R.C., Colli, C.M., Romera, L.I.L. et al. Comparative analysis of routine parasitological methods for recovery of cysts, molecular detection, and genotyping of Giardia duodenalis. Eur J Clin Microbiol Infect Dis 40, 2633–2638 (2021). https://doi.org/10.1007/s10096-021-04280-9

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  • DOI: https://doi.org/10.1007/s10096-021-04280-9

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