Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2005-02-15
2005-02-15
Fredman, Jeffrey (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C536S023100, C536S024300, C536S024320, C435S947000
Reexamination Certificate
active
06855498
ABSTRACT:
The present invention provides in situ hybridization probes which include a marker and a nucleic acid molecule able to hybridize exclusively with only one species ofEncephalitozoon. The nucleic acid molecule may be, for example, complimentary to segment 878-896 of 16S rRNA ofEncephalitozoon hellemspores. Specifically disclosed probes are those including the following nucleotides: (1) 5′-ACT CTC ACA CTC ACT TCA G-3′ (Seq. I.D. No. 1) which is species specific forEncephalitozoon hellem, (2) 5′-CAG ACC ACT ATC TGC A-3′ (Seq. I.D. No. 2) which is species specific forEncephalitozoon cuniculiand (3) 5′-GTT CTC CTG CCC GCT TCA G-3′ (Seq. I.D. No. 3) which is species specific forEncephalitozoon intestinalis. The assay of the present invention utilizes a sample such as surface, ground or drinking water, suspected of containing one of the aforementioned species as a target organism. The microorganisms contained in the sample are fixed in a conventional manner and the probe is then introduced wherein it specifically binds with the target microorganism, if present. The sample is then washed to remove the unbound probe and the bound probe is detected in a conventional manner appropriate for the marker molecule of the probe.
REFERENCES:
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Didier, E. S. et al., “Diagnosis of Disseminated Microsporidian Encephalitozoon hellem Infections by PCR-Southern Analysis and Successful Treatment with Albendazole and Fumagillin”, J. Clin. Microbiol., vol. 34, pp. 947-952 (1996).*
Hester, J. D. et al., “Fluorescent in situ detection of Encephalitozoon hellem spores with a 6-carboxyfluorescein-labeled ribosomal RNA-targeted oligonucleotide probe”, J. Eukaryot. Microbiol., vol. 47, pp. 299-308 ( May-Jun. 2000).
Hester Jeffery Dean
Lindquist H. D. Alan
Schaefer, III Frank W.
Fredman Jeffrey
Lorusso, Loud & Kelly
Loud George A.
Strzelecka Teresa
U.S. Environmental Protection Agency
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