P450 single nucleotide polymorphism biochip analysis

• US 6,986,992 B2
• Filed: 04/01/2002
• Issued: 01/17/2006
• Est. Priority Date: 03/30/2001
• Status: Expired due to Fees

• Current Assignee: Amersham Biosciences (General Electric Company)
• Original Assignee: Amersham Biosciences (General Electric Company)
• Inventors: Luehrsen, Kenneth R., Elghanian, Robert, Mazumder, Abhijit, Yamashiro, Carl, Li, Changming, Gupta, Vineet, Ramakrishnan, Ramesh, Liu, Chang-Gong, Jayaraman, Krishnamurthy, Wang, David G., Yowanto, Handy, Kiser, Gretchen, Chui, Buena, Gu, Zhijie John, Tuggle, Todd, Fermin, David R., Silbergleyt, Arkadiy, Pestova, Ekaterina
• Primary Examiner(s): Switzer, Juliet C.
• Application Number:
  US10/114,908
• Publication Number:
  US 20040229222A1
• Time in Patent Office: 1,387 Days
• Field of Search: 435/6, 435/91.2
• US Class Current: 435/6.1
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2535/125 Allele specific primer extension
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/172 Haplotypes

Abstract

This invention relates to methods and compositions for determining single nucleotide polymorphisms (SNPs) in P450 genes. In preferred embodiments, self extension of interrogation probes is prevented by using novel non self-extension probes and/or methods, thereby improving the specificity and efficiency of P450 SNP detection in target samples with minimal false positive results. The invention thus describes a variety of methods to decrease self-extension of interrogation probes. In addition, this invention provides a unique collection of P450 SNP probes on one assay, primer sequences for specific amplification of each of the seven P450 genes and amplicon control probes to evaluate whether the intended p450 gene targets were amplified successfully. The invention also describes a variety of array platforms for performing the assays of the invention; for example: CodeLink™, eSensor™, multiplex arrays with cartridges etc., all described herein.

Claims

1. A method of determining the identification of a nucleotide at a detection position in a target DNA sequence comprising:

a) amplifying the target DNA using random primers to generate DNA amplicons;

b) transcribing said DNA amplicons to generate RNA target sequences;

c) providing a solid support with a first surface comprising at least one extension probe wherein said extension probe includes an interrogation nucleotide;

d) hybridizing said RNA target sequence to said extension probe to form a hybridization complex;

e) contacting said surface with;

i) a modified reverse transcriptase; and

ii) at least one chain terminating nucleotide comprising a hapten;

under conditions whereby if said chain terminating nucleotide is perfectly complementary to the base of the target sequence immediately adjacent to the 3′

end of extension probe in the hybridization complex, said chain terminating nucleotide is added to extension probe to form a modified extension probe;

f) contacting said modified extension probe with the binding partner of said hapten, wherein said binding partner is labeled; and

g) detecting the presence of said label to determine the nucleotide at said detection position.

2. The method of claim 1, wherein said modified reverse transcriptase only extends extension probes bound to RNA.