Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1999-05-18
2000-12-26
Myers, Carla J.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 405, 435 4051, 536 2432, C12Q 168, G01N 130, C07H 2104
Patent
active
061657239
ABSTRACT:
The present invention provides a method for detecting a target nucleic acid fragment directly from a specimen obtained from a patient by in situ hybridization. The method is comprised of several steps which are performed in the listed order. A sample of the specimen is deposited onto a slide. The sample is fixed onto the slide with fixative, the fixative comprising either methanol-acetic acid at a ratio of from 99:1 to 80:20, or formalin-acetic acid at a ratio of from 99:1 to 80:20. The nucleic acids of the fixed sample are contacted with a probe complex specific for the target nucleic acid fragment, under conditions appropriate for hybridization. Non-hybridized probe complex is rinsed from the sample. The rinsed sample is stained with Evans Blue. The hybridized probe complex is visually detected by microscopy, with the presence of the probe complex being an indication of the presence of the target nucleic acid fragment. The method can be performed with different hybridization buffers, several of which are disclosed. The method of the present invention is useful for detecting pathogens in a specimen. Specific probe complexes are disclosed which are useful for detecting pathogens of the species Babesia. The method is useful in detecting nucleic acids from a wide variety of specimens, including serum, plasma, sputum, urine, cerebral spinal fluids, tissue, breast milk, and insects such as ticks.
REFERENCES:
patent: 4886741 (1989-12-01), Schwartz
patent: 5447841 (1995-09-01), Gray et al.
patent: 5506098 (1996-04-01), Zarling et al.
patent: 5521061 (1996-05-01), Bresser et al.
patent: 5582982 (1996-12-01), Cubbage et al.
patent: 5582985 (1996-12-01), Thompson
patent: 5750340 (1998-05-01), Kim et al.
patent: 5888733 (1999-03-01), Hyldig-Nielsen et al.
Holland, M.S. et al. J. Histochem. Cytochem. 44(3):259-265, Mar. 3, 1996.
DeLong and Shah, Diag. Clin. Testing 28: 41-44 (1990).
DeLong et al., Science 243: 1360-1363 (1989).
Frischner et al., Can. J. Microb. 42: 1061-1071 (1996).
Krause et al., J. Clin. Microbiol. 34: 2014-2016 (1996).
Leidtke et al., PCR Methods and Applications 3: 301-304 (1994).
Shah et al., Clin. Diag. Lab. Immunol. 3: 119-127 (1996).
Amann et al., J. Bacteriol. 172: 762-770 (1990).
Erlandsen et al., (1993) Molecular approach to the speciation and detection of Giardia: fluorochrome-rDNA probes for identification of Giardia lamblia, Giardia Muris, and Giardia ardeae in laboratory and environmental samples by in-situ hybridization. In Giardia --from Molecules to Disease and Beyond, CAB International. pp 64-66.
Roller et al., Microbiology 140: 2849-2858 (1994).
Harris Nick S.
Shah Jyotsna S.
Farrell Kevin M.
Igenex, Inc.
Johannsen Diana
Myers Carla J.
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