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High-Throughput Multiplex Quantitative Polymerase Chain Reaction Method for Giardia lamblia and Cryptosporidium Species Detection in Stool Samples

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  • Department of Virology, School of Medicine, University of Tampere, Finland; Department for International Health, School of Medicine, University of Tampere, Finland; School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi; Department of Pediatrics, Tampere University Hospital, Tampere, Finland; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland

Giardia lamblia and Cryptosporidium species belong to a complex group of pathogens that cause diseases hampering development and socioeconomic improvements in the developing countries. Both pathogens are recognized as significant causes of diarrhea and nutritional disorders. However, further studies are needed to clarify the role of parasitic infections, especially asymptomatic infections in malnutrition and stunting. We developed a high-throughput multiplex quantitative polymerase chain reaction (qPCR) method for G. lamblia and Cryptosporidium spp. detection in stool samples. The sensitivity and specificity of the method were ensured by analyzing confirmed positive samples acquired from diagnostics laboratories and participating in an external quality control round. Its capability to detect asymptomatic G. lamblia and Cryptosporidium spp. infections was confirmed by analyzing stool samples collected from 44 asymptomatic 6-month-old infants living in an endemic region in Malawi. Of these, five samples were found to be positive for G. lamblia and two for Cryptosporidium spp. In conclusion, the developed method is suitable for large-scale studies evaluating the occurrence of G. lamblia and Cryptosporidium spp. in endemic regions and for clinical diagnostics of these infections.

Author Notes

* Address correspondence to Heikki Hyöty, Department of Virology, School of Medicine, University of Tampere, Biokatu 10, FI-33520 Tampere, Finland. E-mail: heikki.hyoty@uta.fi

Financial support: This study was financially supported by Academy of Finland (grants 200720, 108873, 111685, and 109796 [Per Ashorn]), Foundation for Pediatric Research in Finland, Nutriset Inc. (Malaunay, France), Päivikki and Sakari Sohlberg Foundation (Heikki Hyöty), Tampere Tuberculosis foundation (Heikki Hyöty). In addition, part of the study was funded by the DIABIMMUNE project (European Seventh Framework Programme HEALTH-F2-202063).

Authors' addresses: Noora Nurminen, Sami Oikarinen, and Heikki Hyöty, Department of Virology, School of Medicine, University of Tampere, Tampere, Finland, E-mails: noora.nurminen@uta.fi, sami.oikarinen@uta.fi, and heikki.hyoty@uta.fi. Rosa Juuti, EPID Research Oy, Espoo, Finland, E-mail: rosa.mattila@epidresearch.com. Yue-Mei Fan, Kirsi-Maarit Lehto, and Per Ashorn, Department for International Health, School of Medicine, University of Tampere, Tampere, Finland, E-mails: yuemei.fan@uta.fi, kirsi-maarit.lehto@uta.fi, and per.ashorn@uta.fi. Charles Mangani, Department for International Health, School of Medicine, University of Tampere, Tampere, Finland, and College of Medicine, University of Malawi, School of Public Health and Family Medicine, Blantyre, Malawi, E-mail: cmangani@medcol.mw. Kenneth Maleta, School of Public Health and Family Medicine, College of Medicine, University of Malawi, Blantyre, Malawi, E-mail: kmaleta@medcol.mw.

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