Method of amplification for increasing the sensitivity of detect

Details Method of amplification for increasing the sensitivity of detect Method of amplification for increasing the sensitivity of detect

1994-07-15

1997-05-06

Jones, W. Gary

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 5, 536 243, 935 77, 935 88, C07H 2104, C12Q 168

Patent

active

056270305

ABSTRACT:
The present invention provides a method for simplifying and significantly enhancing the sensitivity of nucleic acid hybridization assays. A method is described whereby a single-stranded primary nucleic acid sequence that includes a region of sequences complementary to a single-stranded target nucleic acid sequence is hybridized to the target molecule. Stability of the double-stranded complex thereby formed can be enhanced by using RNA as the probe if DNA is the target or DNA as the probe if RNA is the target. The probe-target complex is subsequently immunocaptured for detection. After washing away extraneous material, a secondary nucleic acid sequence containing many repeating sequence units is hybridized to the probe component of the immobilized probe-target complex. Detection occurs following hybridization of many labelled nucleic acid sequence probes to each of the repeating sequence units of a nucleic acid amplification probe. Thus, attachment of multiple labelling probes to an amplification probe that is hybridized to an immobilized probe-target complex, provides a simplified method for amplifying the detection signal and therefore the sensitivity of nucleic acid hybridization assays.

REFERENCES:
patent: 5124246 (1992-06-01), Urdea et al.
patent: 5175270 (1992-12-01), Nilsen et al.
patent: 5200313 (1993-04-01), Carrico
Matthews, J. et al. Aml. Biochem. 169: 1-25, 1988.

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