Abbott Laboratories Corporation

Hsiang-Yun Y Hu

Daniel Feulner Heiman

John J Carrino

Larry G Birkenmeyer

Jon D Kratochvil

Ronald L Marshall

Natalie A Solomon

John A Walling

Bruce J Dille

Thomas G Laffler

Laurie A Rinehardt

Sharon A Johnson

Barbara E Hasz

An improved, 'gap filling' embodiment of the Ligase Chain Reaction (LCR) is described. Gap filling LCR is LCR wherein at least one of the probes is recessed so that a gap is formed between the adjacent probes when they are hybridized to target. The gap is filled using polymerase and deoxyribonucleotide triphosphates before ligation of the probes together. There are single and double gap versions, depending on whether one or two probes are recessed and require filling before ligation. The improvement resides in selecting and using target sequences such that only a single type, or two types, of deoxyribonucleotide triphosphate(s) are required to fill double gaps each being 1-10 bases in length, preferably 1-3 bases. Probes having specific sequences are claimed for a number of pathogens.

Amplification of target nucleic acids using gap filling ligase chain

This disclosure relates to a method and reagents for determining amphetamine and d-methamphetamine in a biological fluid, such as urine. In particular, this disclosure relates to improvements in a fluorescence polarization immunoassay procedure for determining the presence of amphetamine and d-methamphetamine in a single assay and to a novel class of tracer compounds employed as reagents in such procedures. The procedure described includes pretreatment of the biological sample to eliminate cross reactants such as .beta.-hydroxyphenethylamine by preincubating the sample solely with an aqueous periodate solution having a pH from about 4.0 to about 7.5 without adjustment to an alkaline pH, and contacting the sample with riboflavin binding protein to reduce interference from fluorescent components in the sample. The procedure also maintains the cross reactivity of the immunoassay for tyramine at about 0.4% and for 1-methamphetamine below about 5.1% and eliminates the necessity of using controlled substances as starting materials.

Method and reagents for detecting amphetamine and/or D-methamphetamine

Disclosed is a substantially optically pure hapten, useful in an immunoassay for cotinine. The hapten corresponds to a specified structural formula (I). In formula (I) X is a straight or branched chain, saturated or unsaturated, divalent radical covalently bonded to a pyridyl ring at the number 2-, 4-, 5- or 6-position. The divalent radical, X, has from 1 to 10 carbon atoms wherein the chain of the divalent radical optionally may contain 1 or 2 hetero atoms selected from members of the group consisting of S, O and NZ in which Z represents a C.sub.1 to C.sub.3 alkyl group. In formula (I), n=1 or 0, and Q is a functional group selected from --COOH, --NH.sub.2 (provided that n does not equal 0), --C(O)NHNH.sub.2, --O(CO)Cl, --CHO, --NCS or --NCO. Also disclosed is an immunogen derived from the hapten as well as an antibody raised in response to an immunogen derived from the hapten. Also disclosed is a fluorescent tracer derived from a substantially optically pure compound corresponding to the hapten, the tracer being useful in an immunoassay for cotinine. Also disclosed is an improved immunoassay for determining cotinine in a biological sample involving a step of contacting the sample with antibodies raised in response to the immunogen. Also disclosed is a fluorescence polarization immunoassay (FPIA) for determining cotinine involving a step of contacting the sample with antibodies raised in response to the immunogen, and/or involving a step of contacting the sample with a fluorescent tracer.

Haptens, tracers, immunogens and antibodies for immunoassays for

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Libertyville, IL, United States of America

Hsiang-Yun Y Hu