Giardia and Cryptosporidium spp. dissemination during wastewater treatment and comparative detection via immunofluorescence assay (IFA), nested polymerase chain reaction (nested PCR) and loop mediated isothermal amplification (LAMP)

Carmen Gallas-Lindemann; Isaia Sotiriadou; Judit Plutzer; Michael J Noack; Mohammad Reza Mahmoudi; Panagiotis Karanis

doi:10.1016/j.actatropica.2016.02.005

Giardia and Cryptosporidium spp. dissemination during wastewater treatment and comparative detection via immunofluorescence assay (IFA), nested polymerase chain reaction (nested PCR) and loop mediated isothermal amplification (LAMP)

Acta Trop. 2016 Jun:158:43-51. doi: 10.1016/j.actatropica.2016.02.005. Epub 2016 Feb 12.

Authors

Carmen Gallas-Lindemann¹, Isaia Sotiriadou², Judit Plutzer³, Michael J Noack⁴, Mohammad Reza Mahmoudi⁵, Panagiotis Karanis⁶

Affiliations

¹ Medical and Molecular Parasitology Laboratory, University of Cologne, Medical School, Center of Anatomy, Institute II, Cologne, Germany; Linksniederrheinische Entwässerungs-Genossenschaft, Friedrich-Heinrich-Allee 64, 47475, Kamp-Lintfort, Germany. Electronic address: gallas-lindemann.c@lineg.de.
² Medical and Molecular Parasitology Laboratory, University of Cologne, Medical School, Center of Anatomy, Institute II, Cologne, Germany; Centre of Dental Medicine, Policlinic of Operative Dentistry and Periodontology, University of Cologne, Germany.
³ National Public Health Center, National Directorate of Environmental Health, Environmental Health Testing Laboratory, Budapest, Hungary.
⁴ Centre of Dental Medicine, Policlinic of Operative Dentistry and Periodontology, University of Cologne, Germany.
⁵ Department of Medical Microbiology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
⁶ Medical and Molecular Parasitology Laboratory, University of Cologne, Medical School, Center of Anatomy, Institute II, Cologne, Germany; Center Biomedicine and Infectious Diseases (CBID), One Thousand Talent Program of the Chinese Government, Qinghai Academy for Veterinary Medicine and Animal Sciences, Qinghai University, Xining City, 1# Wei'er Road, Qinghai Biological Scientific Estate Graden, Xining City, Qinghai Province 810016, PR China.

PMID: 26880717
DOI: 10.1016/j.actatropica.2016.02.005

Abstract

Environmental water samples from the Lower Rhine area in Germany were investigated via immunofluorescence assays (IFAs), nested polymerase chain reaction (nested PCR) and loop-mediated isothermal amplification (LAMP) to detect the presence of Giardia spp. (n=185) and Cryptosporidium spp. (n=227). The samples were concentrated through filtration or flocculation, and oocysts were purified via centrifugation through a sucrose density gradient. For all samples, IFA was performed first, followed by DNA extraction for the nested PCR and LAMP assays. Giardia cysts were detected in 105 samples (56.8%) by IFA, 62 samples (33.5%) by nested PCR and 79 samples (42.7%) by LAMP. Cryptosporidium spp. were detected in 69 samples (30.4%) by IFA, 95 samples (41.9%) by nested PCR and 99 samples (43.6%) by LAMP. According to these results, the three detection methods are complementary for monitoring Giardia and Cryptosporidium in environmental waters.

Keywords: Cryptosporidium; Detection; Dissemination; Giardia; LAMP; Wastewater.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cryptosporidium / genetics*
Cryptosporidium / isolation & purification*
Fluorescent Antibody Technique, Direct
Germany
Giardia / genetics*
Giardia / isolation & purification*
Nucleic Acid Amplification Techniques
Polymerase Chain Reaction
Wastewater / parasitology*

Substances

Waste Water