Method for obtaining purified RNA from viable oocysts
First Claim
Patent Images
1. A method for obtaining purified RNA from viable oocysts, the method comprising the steps of:
- a) centrifuging a fluid sample containing oocysts at a speed and for a period of time sufficient to concentrate the oocysts within a vessel containing the fluid sample;
b) removing a supernatant formed in the vessel during the centrifuging step;
c) resuspending the concentrated oocysts in a buffered solution;
d) agitating the buffered solution in the presence of zirconia/silica particles at a rate and for a period of time sufficient to lyse the oocysts and release RNA therefrom;
e) immobilizing the released RNA on an RNA-binding filter;
f) purifying the released RNA by washing the filter one or more times with a buffered solution to remove oocyst components other than the released RNA; and
g) removing the purified RNA from the filter.
5 Assignments
0 Petitions
Accused Products
Abstract
The present invention describes novel oligonucleotides targeted to nucleic acid sequences derived from Crytosporidium organisms, and Crytosporidium parvum organisms in particular, which are useful for determining the presence of Cryptosporidium organisms in a test sample. The oligonucleotides of the present invention include hybridization assay probes, helper probes and amplification primers. The present invention further describes a novel method for obtaining purified ribonucleic acid from viable oocysts.
36 Citations
14 Claims
-
1. A method for obtaining purified RNA from viable oocysts, the method comprising the steps of:
-
a) centrifuging a fluid sample containing oocysts at a speed and for a period of time sufficient to concentrate the oocysts within a vessel containing the fluid sample; b) removing a supernatant formed in the vessel during the centrifuging step; c) resuspending the concentrated oocysts in a buffered solution; d) agitating the buffered solution in the presence of zirconia/silica particles at a rate and for a period of time sufficient to lyse the oocysts and release RNA therefrom; e) immobilizing the released RNA on an RNA-binding filter; f) purifying the released RNA by washing the filter one or more times with a buffered solution to remove oocyst components other than the released RNA; and g) removing the purified RNA from the filter. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
-
Specification