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Comparison of various staining methods for the detection of Cryptosporidium in cell-free culture
Journal article   Peer reviewed

Comparison of various staining methods for the detection of Cryptosporidium in cell-free culture

A. Boxell, N. Hijjawi, P. Monis and U. Ryan
Experimental Parasitology, Vol.120(1), pp.67-72
2008
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Abstract

The complete development of Cryptosporidium in host cell-free medium first described in 2004, represented a significant advance that can facilitate many aspects of Cryptosporidium research. A current limitation of host cell-free cultivation is the difficulty involved in visualising the life-cycle stages as they are very small in size, morphologically difficult to identify and dispersed throughout the media. This is in contrast to conventional cell culture methods for Cryptosporidium, where it is possible to focus on the host cells and view the foci of infection on the host cells. In the present study, we compared three specific and three non-specific techniques for visualising Cryptosporidium parvum life-cycle stages in cell-free culture; antibody staining using anti-sporozoite and anti-oocyst wall antibodies (Sporo-Glo™ and Crypto Cel), fluorescent in-situ hybridization (FISH) using a Cryptosporidium specific rRNA oligonucleotide probe and the non-specific dyes; Texas Red, carboxyfluorescein diacetate succinimidyl ester (CFSE) and 4,6′ diamino-2-phenylindole dihydrochloride (DAPI). Results revealed that a combination of Sporo-Glo™ and Crypto Cel staining resulted in easy and reliable identification of all life-cycle stages.

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Collaboration types
Domestic collaboration
International collaboration
Citation topics
1 Clinical & Life Sciences
1.246 Diarrheal Diseases
1.246.985 Cryptosporidium
Web Of Science research areas
Parasitology
ESI research areas
Microbiology
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