A novel nested multiplex PCR for the simultaneous detection and differentiation of Cryptosporidium spp., Enterocytozoon bieneusi and Encephalitozoon intestinalis

The development of molecular methodologies for targeting pathogens such as the Microsporidia has greatly improved our monitoring capabilities and initiatives. This study analyzed samples collected from five locations in Pensacola, Florida, USA for the presence of Microsporidian pathogens. To circumvent various impediments associated with water collection and filtration, we utilized biofilms as sentinels for detection of Microsporidia. We implemented membrane-dissolution and sample purification in a single confined step followed by real-time PCR to confirm pathogen presence. The results of this study demonstrate that microsporidia are present in environmental water sites in the Florida panhandle and that biofilms may serve as another alternative mode to circumvent filtration methods for their detection.

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“…Species‐specific primers for E. cuniculi , E. intestinalis and E. hellem have been described (Rubio et al. ; Valencakova et al. ).…”

Section: Discussionmentioning

confidence: 99%

“…A follow-up study to ascertain which Encephalitozoon species are routinely present would be ideal. Species-specific primers for E. cuniculi, E. intestinalis and E. hellem have been described (Rubio et al 2014;Valencakova et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Biofilms for Monitoring Presence of Microsporidia in Environmental Water

Moss

Snyder²

2017

J Eukaryotic Microbiology

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“…Cryptosporidiosis, caused by the enteric parasite pathogens Cryptosporidium spp., can lead to diarrheal illness in humans and animals including birds [ 1 – 3 ]. Since Tyzzer [ 4 ] firstly observed the Cryptosporidium infection in birds, this pathogen has been detected in more than 30 avian species worldwide [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

Prevalence and Genotyping ofCryptosporidiumInfection in Pet Parrots in North China

Zhang

Zhao

et al. 2015

BioMed Research International

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Cryptosporidiosis is a worldwide zoonosis caused by Cryptosporidium spp., sometimes leading to severe diarrhea in humans and animals. In the present study, 311 parrots, belonging to four species, namely, Budgerigars (Melopsittacus undulatus), Lovebirds (Agapornis sp.), Alexandrine parakeets (Psittacula eupatria), and Cockatiel (Nymphicus hollandicus), from Beijing and Weifang cities, were examined for Cryptosporidium spp. infection. Blood samples of each bird were examined using enzyme linked immunosorbent assay (ELISA) and fecal samples were examined by Sheather's sugar flotation technique. Prevalence of Cryptosporidium infection were 3.22% (10/311) and 0.64% (2/311) by ELISA and Sheather's sugar flotation technique, respectively. Seroprevalence of Cryptosporidium infection in different breeds varied from 0 to 15.39%. Sequencing analysis showed that both positive samples from fecal samples belonged to Cryptosporidium avian genotype V. This is the first report of Cryptosporidium avian genotype V in Budgerigars. The results of the present study provided foundation-data for prevention and control of cryptosporidiosis in pet birds in China.

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“…Overall, several studies have recommended the incorporation of PCR techniques into routine clinical Cryptosporidium diagnosis methods ( Rubio et al, 2014 ; Stensvold et al, 2011 ; Uppal et al, 2014 ). Indeed, PCR has been reported as having improved sensitivity over current detection methods, with reported limits of detection ranging from 1 × 10 5 to 1 oocyst/Gram of faeces ( Costa et al, 2021 ).…”

Section: Diagnosismentioning

confidence: 99%

Cryptosporidium spp. Diagnosis and Research in the 21st Century

O’Leary

Sleator

Lucey

2021

Food and Waterborne Parasitology

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The protozoan parasite Cryptosporidium has emerged as a leading cause of diarrhoeal illness worldwide, posing a significant threat to young children and immunocompromised patients. While endemic in the vast majority of developing countries, Cryptosporidium also has the potential to cause waterborne epidemics and large scale outbreaks in both developing and developed nations. Anthroponontic and zoonotic transmission routes are well defined, with the ingestion of faecally contaminated food and water supplies a common source of infection. Microscopy, the current diagnostic mainstay, is considered by many to be suboptimal. This has prompted a shift towards alternative diagnostic techniques in the advent of the molecular era. Molecular methods, particularly PCR, are gaining traction in a diagnostic capacity over microscopy in the diagnosis of cryptosporidiosis, given the laborious and often tedious nature of the latter. Until now, developments in the field of Cryptosporidium detection and research have been somewhat hampered by the intractable nature of this parasite. However, recent advances in the field have taken the tentative first steps towards bringing Cryptosporidium research into the 21 st century. Herein, we provide a review of these advances.

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A novel nested multiplex PCR for the simultaneous detection and differentiation of Cryptosporidium spp., Enterocytozoon bieneusi and Encephalitozoon intestinalis

Cited by 13 publications

References 36 publications

Biofilms for Monitoring Presence of Microsporidia in Environmental Water

Biofilms for Monitoring Presence of Microsporidia in Environmental Water

Prevalence and Genotyping ofCryptosporidiumInfection in Pet Parrots in North China

Cryptosporidium spp. Diagnosis and Research in the 21st Century

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