PMID- 9750273
OWN - NLM
STAT- MEDLINE
DCOM- 19981207
LR  - 20220417
IS  - 1364-5072 (Print)
IS  - 1364-5072 (Linking)
VI  - 85
IP  - 3
DP  - 1998 Sep
TI  - The use of a ribosomal RNA targeted oligonucleotide probe for fluorescent 
      labelling of viable Cryptosporidium parvum oocysts.
PG  - 429-40
AB  - A fluorescence in situ hybridization (FISH) technique has been developed for the 
      fluorescent labelling of Cryptosporidium parvum oocysts in water samples. The 
      FISH technique employs a fluorescently labelled oligonucleotide probe (Cry1 
      probe) targeting a specific sequence in the 18S ribosomal RNA (rRNA) of C. 
      parvum. Hybridization with the Cry1 probe resulted in fluorescence of sporozoites 
      within oocysts that were capable of excystation, while oocysts that were dead 
      prior to fixation did not fluoresce. Correlation of the FISH method with 
      viability as measured by in vitro excystation was statistically highly 
      significant, with a calculated correlation coefficient of 0.998. Examination of 
      sequence data for Cryptosporidium spp. other than C. parvum suggests that the 
      Cry1 probe is C. parvum-specific. In addition, 19 isolates of C. parvum were 
      tested, and all fluoresced after hybridization with the Cry1 probe. Conversely, 
      isolates of C. baileyi and C. muris were tested and found not to fluoresce after 
      hybridization with the Cry1 probe. The fluorescence of FISH-stained oocysts was 
      not bright enough to enable detection of oocysts in environmental water 
      concentrates containing autofluorescent algae and mineral particles. However, in 
      combination with immunofluorescence staining, FISH enabled species-specific 
      detection and viability determination of C. parvum oocysts in water samples.
FAU - Vesey, G
AU  - Vesey G
AD  - Macquarie University Centre for Analytical Biotechnology, School of Biological 
      Sciences, Macquarie University, New South Wales, Australia. 
      gvesey@rna.bio.mq.edu.au
FAU - Ashbolt, N
AU  - Ashbolt N
FAU - Fricker, E J
AU  - Fricker EJ
FAU - Deere, D
AU  - Deere D
FAU - Williams, K L
AU  - Williams KL
FAU - Veal, D A
AU  - Veal DA
FAU - Dorsch, M
AU  - Dorsch M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Appl Microbiol
JT  - Journal of applied microbiology
JID - 9706280
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 18S)
RN  - I223NX31W9 (Fluorescein-5-isothiocyanate)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/immunology
MH  - Cell Membrane Permeability
MH  - Cryptosporidium parvum/genetics/*isolation & purification
MH  - Fluorescein-5-isothiocyanate
MH  - Fluorescent Dyes
MH  - Gene Amplification
MH  - In Situ Hybridization, Fluorescence/*methods/standards
MH  - *Oligonucleotide Probes
MH  - RNA, Bacterial/analysis
MH  - RNA, Ribosomal, 18S/analysis
MH  - Sensitivity and Specificity
MH  - Staining and Labeling
MH  - Water Supply
EDAT- 1998/09/29 00:00
MHDA- 1998/09/29 00:01
CRDT- 1998/09/29 00:00
PHST- 1998/09/29 00:00 [pubmed]
PHST- 1998/09/29 00:01 [medline]
PHST- 1998/09/29 00:00 [entrez]
AID - 10.1046/j.1365-2672.1998.853496.x [doi]
PST - ppublish
SO  - J Appl Microbiol. 1998 Sep;85(3):429-40. doi: 10.1046/j.1365-2672.1998.853496.x.