Cryptosporidium spp. Identification, Quantification and Genotyping

Identification, quantification and genotyping of Cryptosporidium spp.

Human stool sample containing Cryptosporidium

Human stool sample may be fresh or in transport media such as Cary-Blair. Samples in fixatives containing formalin, such as SAF are not accepted.

Samples contained in a leak-proof container with sample number on the tube and any identifying patient information removed. Preferred minimum sample size is 1 gram. Samples may be frozen on dry ice or refrigerated on wet ice. Repeated freeze-thaws should be avoided if possible.

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

Watery diarrhea, fatigue, anorexia, myalgias, weight loss, abdominal pain, low-grade fever, nausea and vomiting.

Completed Parasite Genomics Unit requisition form.

Samples in fixatives containing formalin, such as SAF, are not accepted.

Molecular characterization by PCR and sequencing (1)

Molecular quantification by qPCR (1, 2, 3)

Genotyping using CryptoGenotyper bioinformatics tool (4) for species identification and gp60 subtype.

1. Molecular genotyping method as per: Guy RA, Yanta CA, Muchaal PK, et al. Molecular characterization of Cryptosporidium isolates from humans in Ontario, Canada. Parasites Vectors (2021) 14 69. doi: 10.1186/s13071-020-04546-9
2. qPCR - C. parvum and C. hominis as per: Robinson G, Chalmers RM. Assessment of polymorphic genetic markers for multi-locus typing of Cryptosporidium parvum and Cryptosporidium hominis. Experimental Parasitology 2012 132:200–215. doi: 10.1016/j.exppara.2012.06.016.
3. qPCR - Cryptosporidium spp. modified from: Stroup SE, Roy S, Mchele J, Maro V, Ntabaguzi S, Siddique A, Kang G, Guerrant RL, Kirkpatrick BD, Fayer R, Herbein J, Ward H, Haque R, Houpt ER. Real-time PCR detection and speciation of Cryptosporidium infection using Scorpion probes. J Med Microbiol. 2006 Sep;55(Pt 9):1217-1222. doi: 10.1099/jmm.0.46678-0.
4. In silico genotyping as per: Yanta CA, Bessonov K, Robinson G, Troell K, Guy RA. CryptoGenotyper: A new bioinformatics tool for rapid Cryptosporidium identification. Food Waterborne Parasitol. 2021 Feb 26;23:e00115. doi: 10.1016/j.fawpar.2021.e00115.

7 calendar days. Note: Turnaround times for routine isolates may be extended during outbreak activities or due to limited availability of resources.