Genotyping Cryptosporidium parvum with an hsp70 single-nucleotide polymorphism microarray.

Applied and Environmental Microbiology
T M StraubDarrell P Chandler

Abstract

We investigated the application of an oligonucleotide microarray to (i) specifically detect Cryptosporidium spp., (ii) differentiate between closely related C. parvum isolates and Cryptosporidium species, and (iii) differentiate between principle genotypes known to infect humans. A microarray of 68 capture probes targeting seven single-nucleotide polymorphisms (SNPs) within a 190-bp region of the hsp70 gene of Cryptosporidium parvum was constructed. Labeled hsp70 targets were generated by PCR with biotin- or Cy3-labeled primers. Hybridization conditions were optimized for hybridization time, temperature, and salt concentration. Two genotype I C. parvum isolates (TU502 and UG502), two C. parvum genotype II isolates (Iowa and GCH1), and DNAs from 22 non-Cryptosporidium sp. organisms were used to test method specificity. Only DNAs from C. parvum isolates produced labeled amplicons that could be hybridized to and detected on the array. Hybridization patterns between genotypes were visually distinct, but identification of SNPs required statistical analysis of the signal intensity data. The results indicated that correct mismatch discrimination could be achieved for all seven SNPs for the UG502 isolate, five of seven SNPs for the TU5...Continue Reading

References

Sep 1, 1990·DNA and Cell Biology·R DrmanacR Crkvenjakov
Jun 1, 1995·The American Journal of Tropical Medicine and Hygiene·U M MorganR C Thompson
May 24, 1994·Proceedings of the National Academy of Sciences of the United States of America·A C PeaseS P Fodor
May 14, 1996·Proceedings of the National Academy of Sciences of the United States of America·G YershovA Mirzabekov
Jun 1, 1997·Applied and Environmental Microbiology·D Y GuschinA D Mirzabekov
Dec 1, 1996·Nature Biotechnology·D J LockhartE L Brown
Apr 2, 1999·Applied and Environmental Microbiology·L XiaoA A Lal
Oct 9, 1999·Clinical Microbiology Reviews·D P Clark
Jan 5, 2000·International Journal for Parasitology·U M MorganR C Thompson
Jun 6, 2000·Molecular and Biochemical Parasitology·G WidmerS Tzipori
Aug 5, 2000·Applied and Environmental Microbiology·X FengG Widmer
Mar 10, 2001·BioTechniques·D R CallF Brockman
Mar 27, 2001·Emerging Infectious Diseases·S Pedraza-DíazJ McLauchlin
May 31, 2001·Journal of Computational Biology : a Journal of Computational Molecular Cell Biology·M K KerrG A Churchill
Sep 26, 2001·Applied and Environmental Microbiology·J SmallD P Chandler
Nov 27, 2002·Applied and Environmental Microbiology·Carol L DiGiorgioChristopher C Huitt

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Citations

Mar 26, 2003·Journal of Microbiological Methods·Timothy M Straub, Darrell P Chandler
Jul 2, 2003·Current Opinion in Microbiology·Jizhong Zhou
Dec 31, 2002·The Journal of Eukaryotic Microbiology·Una M Morgan-RyanLihua Xiao
Jun 11, 2004·Eukaryotic Cell·Anja ForcheGeorgiana May
Dec 9, 2009·Journal of Microbiological Methods·Dae-Young LeeRenata Korczak
Nov 22, 2012·Journal of Applied Microbiology·N E BrinkmanE N Villegas
Nov 4, 2004·Biosensors & Bioelectronics·Samantha LaneDouglas R Call
Oct 24, 2006·Biotechnology Advances·Jeanine I Boulter-BitzerJack T Trevors
Aug 30, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Alexander Loy, Levente Bodrossy
Oct 20, 2004·Critical Reviews in Microbiology·Karine LemarchandRoland Brousseau
Mar 30, 2004·Biopolymers·Arnold Vainrub, B Montgomery Pettitt
Jul 2, 2005·Critical Reviews in Microbiology·Douglas R Call

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