FAQ
Jagiellonian University logo

New primers for fast detection of Giardia duodenalis assemblages A and B using realtime PCR

Publication date: 19.09.2018

Acta Protozoologica, 2018, Volume 57, Issue 1, pp. 43 - 48

https://doi.org/10.4467/16890027AP.18.003.8397

Authors

,
Piotr Solarczyk
Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland
https://orcid.org/0000-0002-2862-4742 Orcid
All publications →
,
Agnieszka Wojtkowiak-Giera
Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland
All publications →
,
Marcin Hołysz
Department of Biochemistry and Molecular Biology, Poznan University of Medical Sciences, Poznan, Poland
All publications →
,
Anna Słodkowicz-Kowalska
Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland
All publications →
,
Paweł P. Jagodziński
Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland
All publications →
,
Krzysztof Stojecki
Department of Parasitology, National Veterinary Research Institute, Pulawy, Poland
All publications →
,
Anna Rocka
The Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Warszawa, Poland
All publications →
,
Anna C. Majewska
Poznan University of Medical Sciences
, Poland
All publications →
Łukasz Skrzypczak
Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland
All publications →

Titles

New primers for fast detection of Giardia duodenalis assemblages A and B using realtime PCR

Abstract

iardia duodenalisis one of the six Giardia species and itis the most common, cosmopolitan flagellate that infects humans and many species of animals.This species exhibits considerable genetic diversity; to date, eight assemblages (A–H) have been defined. These assemblages differ in host specificity: assemblages A and B have beenfound in both humans and in many animal species. Mixed infections with Giardia (A and B) assemblages have been reported in humans and in animals. Many molecular techniques are effective and rapid for the detection of G. duodenalis and also forthe determination of genetic variability of isolates in clinical and environmental samples. In this context, the aim of this study was to design new assemblage-specific primers for rapid detection and identification ofG. duodenalis assemblages A and B and both of these assemblages simultaneously using quantitative real-time polymerase chain reaction (qPCR). Fragments of glutamate dehydrogenase and triose phosphate isomerase were used as targets in the design of primers.
In conclusion, the use of G. duodenalis assemblage-specific primers designed in this study allows quick identification of human infectious G. duodenalis assemblages A and B as well as mixed AB assemblages in a sample without further sequencing of the amplification products, which reduces the cost of study and the waiting time for the results.

References

Download references

Almeida A., Pozio E., Cacció S. M. (2010) Genotyping of Giardia duodenalis cysts by new real-time PCR assay for detection of mixed infections in human samples. Appl. Environ. Microbiol. 76: 1895–1901

Alonso J. L., Amoros, Guy R. A. (2014) Quantification of viable Giardia cysts and cryptosporidium oocysts in wastewater using propidium monoazide quantitative real-time PCR. Parasitol. Res. 113: 2671–2267

Amar C. F., Dear, P. H., Pedraza-Diaz S., Looker N., Linnane E., McLauchlin J. (2002) Sensitive PCR-restriction fragment length polymorphism assay for detection and genotyping of Giardia intestinalis in human feces. J. Clin. Microbiol. 40: 446–452

Cacciò S. M., Ryan U. (2008) Molecular epidemiology of giardiasis. Mol. Biochem. Parasitol. 160: 75–80

Feng Y., Xiao L. (2011) Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin. Microbiol. Rev. 24: 110–140

Gotfred-Rasmussen H., Lund M., Enemark H. L., Erlandsen M., Petersen E. (2016) Comparison of sensitivity and specificity of 4 methods for detection of Giardia duodenalis in feces: immunofluorescence and PCR are superior to microscopy of concentrated iodine-stained samples. Diagn. Microbiol. Infect. Dis. 84: 187–190

Gizzi A. B, Oliveira T. S., Leutenegger C. M., Estrada M., Kozemjakin D. A., Stedile R., Marcondes M., Biondo A. W. (2014) Presence of infectious agents and co-infections in diarrheic dogs determined with a real-time polymerase chain reaction-based panel. BMC Vet. Res. 10: 1–8

Guy R. A., Payment P., Ulrich J. K., Horgen P. A. (2003) Real-time PCR for quantification of Giardia and Cryptosporidium in environmental water samples and sewage. Appl. Enviro. Microbiol. 69: 5178–5185

Guy R. A., Xiao Ch., Paul A. H. (2004) Real-time PCR assay for detection and genotype differentiation of Giardia lamblia in stool specimens. J. Clin. Microbiol42: 3317–3320

Heyworth M. F. (2016) Giardia duodenalis genetic assemblages and hosts. Parasite 23: 1–5

Jerlstorm-Hultqvist J., Ankarklev J., Svard S. G. (2010) Is human giardiasis caused by two different Giardia species? Gut Microbes1: 379–382

McGlade T. R., Robertson I. D., Elliot A. D., Read C, Thompson R. C. (2003) Gastrointestinal parasites of domestic cats in Perth, Western Australia. Vet. Parasitol. 117: 251–262

Monis P. T., Caccio S. M., Thompson R. C. (2009) Variation in Giardia: towards a taxonomic revision of the genus. Trends Parasitol. 25: 93–100

Pallant L., Barutzki D., Schaper R., Thompson R. C. A. (2015) The epidemiology of infections with Giardia species and genotypes in well cared for dogs and cats in Germany. Parasit. Vectors 8: 2–14

Prasertbun R., Sukthana Y., Popruk S. (2012) Real-time PCR: Benefits for detection of mild and asymptomatic Giardia infections. Trop. Med. Health 40: 31–35

Ryan U., Cacciò S. M. (2013) Zoonotic potential of GiardiaInt. J. Parasitol. 43: 943–956

Solarczyk P., Werner A., Majewska A. C. (2010) Genotypowanie izolatów Giardia duodenalis uzyskanych od ludzi w zachodnio-centralnej Polsce. Wiad. Parazytol. 56: 71–177

Schuurman T., Lankamp P., van Belkum A., Kooistra-Smid M., van Zwet A. (2007) Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens. Clin. Microbiol. Infect. 13: 1186–1191

Skrzypczak Ł. (2016) Występowanie stadiów dyspersyjnych Giardia i Cryptosporidium w wodzie z fontann i kąpielisk. [Occurrence of dispersive stages of Giardia and Cryptosporidium in water from fountains and swimming sites] Doctoral thesis. Poznań University of Medical Sciences, Department of Biology and Medical Parasitology, 1–109

Solarczyk P., Majewska A. (2010) A survey of the prevalence and genotypes of Giardia duodenalis infecting household and sheltered dogs. Parasitol. Res. 106: 1015–1019

Solarczyk P., Majewska A. C., Moskwa B., Cabaj W., Dabert M., Nowosad P. (2012) Multilocus genotyping of Giardia duodenalis isolates from red deer (Cervus elaphus) and roe deer (Capreolus capreolus) from Poland. Folia Parasitol. (Prahga) 59: 237–240

Solarczyk P., Majewska A. C., Słodkowicz-Kowalska A. (2014) Axenic in vitro culture and molecular characterization of Giardia duodenalis from red deer (Cervus elaphus) and Thomson’s gazelle (Gazella thomsonii). Acta Parasitol. 59: 763–766

Solarczyk P., Majewska A. C., Jędrzejewski S., Górecki M. T., Nowicki S., Przysiecki P. (2016) First record of Giardia assemblage D infection in farmed raccoon dogs (Nyctereutes procyonoides). Ann. Agric. Environ. Med. 23: 595–597

Sprong H., Cacciò S. M., van der Giessen J. W. (2009) ZOOPNET network and partners. Identification of zoonotic genotypes of Giardia duodenalis. PLoS Negl. Trop. Dis. 3: 1–12

Stojecki K., Sroka J., Cencek T., Dutkiewicz J. (2015) Epidemiological survey in Łęczyńsko-Włodawskie Lake District of eastern Poland reveals new evidence of zoonotic potential of Giardia intestinalisAnn. Agric. Environ. Med. 22: 594–598

Xu F., Jerlstorm-Hultqvist J., Andersson J. O. (2012) Genome-wide analyses of recombination suggest that Giardia intestinalis assemblages represent different species. Mol. Biol. Evol. 29: 2859–2858

Zhang P., Liu P., Alsarakibi M., Li J., Liu T., Li Y., Li G. 2012. Application of HRM assays with EvaGreen dye for genotyping Giardia duodenalis zoonotic assemblages. Parasitol. Res. 111: 2157–2163

Zhang H., Zhang X., Zhang S., Wei B., Jiang Q., Yu X. (2013) Detecting Cryptosporidium parvum and Giardia lamblia by coagulation concentration and real-time PCR quantification. F. E. S. E. 7: 49–54

Vanni I., Caccio S. M., van Lith L., Lebbad M., Svard S. G., Pozio E., Tosini F. (2012) Detection of Giardia duodenalis assemblages A and B in human feces by simple, assemblage-specific PCR assays. PLoS Negl. Trop. Dis. 6: 1–9

Verweij J. J., Schinkel J., Laeijendecker D., van Rooyen M. A. A., van Lieshout L., Polderman A. M. (2003) Real-time PCR for the detection of Giardia lambliaMol. Cell. Probes. 17: 223–225

Xiao L., Fayer R. (2008) Molecular characterization of species and genotypes of Giardia and Cryptosporidium and assessment of zoonotic transmission. Int. J. Parasitol. 38: 1239–1255

Information

Information: Acta Protozoologica, 2018, Volume 57, Issue 1, pp. 43 - 48

Article type: Original article

Authors

https://orcid.org/0000-0002-2862-4742

Piotr Solarczyk
Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland
https://orcid.org/0000-0002-2862-4742 Orcid
All publications →

Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland

Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland

Department of Biochemistry and Molecular Biology, Poznan University of Medical Sciences, Poznan, Poland

Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland

Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland

Department of Parasitology, National Veterinary Research Institute, Pulawy, Poland

The Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Warszawa, Poland

Poznan University of Medical Sciences
Poland

Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, Poznan, Poland

Published at: 19.09.2018

Article status: Open

Licence: CC BY-NC-ND  licence icon

Percentage share of authors:

Piotr Solarczyk (Author) - 11%
Agnieszka Wojtkowiak-Giera (Author) - 11%
Marcin Hołysz (Author) - 11%
Anna Słodkowicz-Kowalska (Author) - 11%
Paweł P. Jagodziński (Author) - 11%
Krzysztof Stojecki (Author) - 11%
Anna Rocka (Author) - 11%
Anna C. Majewska (Author) - 11%
Łukasz Skrzypczak (Author) - 12%

Article corrections:

-

Publication languages:

English