Methods and apparatus for preparing, amplifying, and discriminating multiple analytes
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12P-019/34
C07H-021/00
C07H-021/04
출원번호
US-0692726
(1996-08-06)
발명자
/ 주소
Wu Linxian
Coombs Jana
Malmstrom Sharon L.
Glass Michael J.
출원인 / 주소
Gull Laboratories
대리인 / 주소
Workman, Nydegger & Seeley
인용정보
피인용 횟수 :
39인용 특허 :
0
초록▼
The present invention provides methods and apparatus for detecting and discriminating multiple analytes within a test sample which are simple, user-friendly, cost-effective and fast. In particular, it is preferred that the overall time for sample preparation, nucleic acid sequence amplification, and
The present invention provides methods and apparatus for detecting and discriminating multiple analytes within a test sample which are simple, user-friendly, cost-effective and fast. In particular, it is preferred that the overall time for sample preparation, nucleic acid sequence amplification, and nucleic acid sequence differentiation be about 5 hours or less. The methods of the present invention comprise (i) rapid sample processing means for rapidly preparing sample material of various types for amplification of nucleic acid sequences using unique nucleic acid extraction buffer formulations, (ii) multianalyte non-preferential amplifying process means for simultaneously and non-preferentially amplifying multiple target nucleic acid sequences, if present within the sample, using appropriate primer oligonucleotides optimized to achieve substantially similar amplification efficiencies, and (iii) multianalyte recognition process means for detecting and discriminating amplified nucleic acid sequences which incorporate nucleic acid sequence mismatch detection means for differentiating minor mismatches between multiple amplified nucleic acid sequences, including only single base mismatches, using appropriate probe oligonucleotides modified with neutral base substitution molecules. The processing kit products in accord with the present invention may incorporate all, or only some, of the above-described means.
대표청구항▼
[ What is claimed and desired to be secured by United States Letters Patent is:] [1.] A method for simultaneously and non-preferentially amplifying multiple target nucleic acid sequences from multiple microorganisms, if present within a sample containing nucleic acids from multiple microorganisms, s
[ What is claimed and desired to be secured by United States Letters Patent is:] [1.] A method for simultaneously and non-preferentially amplifying multiple target nucleic acid sequences from multiple microorganisms, if present within a sample containing nucleic acids from multiple microorganisms, said multiple target nucleic acid sequences including microorganisms of the Salmonella, Shigella, Campylobacter, Yersinia, and Escherichia coli species, said method comprising the steps of:obtaining multiple primer oligonucleotides, each said primer oligonucleotide being complementary to a portion of one strand of a target nucleic acid sequence and specific for one of said multiple microorganisms;selecting processing conditions for a simultaneous polymerase chain reaction amplification;mixing said sample with said multiple primer oligonucleotides in selected concentrations such that substantially similar amplification efficiencies under the selected processing conditions are achieved for each target nucleic acid sequence present in said sample; andperforming a polymerase chain reaction amplification on said mixture; rein the step of obtaining multiple primer oligonucleotides comprises obtaining SEQ ID NO:1 and SEQ ID NO:2 for use in detecting target nucleic acid sequences from microorganisms of the species Salmonella.
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