TH33558A - The quantification and identification of various species of substances including Myocbacterium avium. - Google Patents

Abstract

The amplification primers and methods for group-specific amplification of Chinese dnaJ target sequences of Mycobacterium species have been revealed. Methods for detecting amplified products and/or identifying various existing MAC species have also been provided.

Claims (10)

1. A bulking primer containing a binding sequence to a selected target from a group of binding sequences SEQ [D NO:1, SEQ ID NO:2, and SEQ ID NO:3]. 2. A primer for increasing the quantity of claim 1, selected from a group consisting of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3. 3. Methods for group-specific amplification of Mycobacterium avium target nucleic acids consist of: a) hybridizing the target nucleic acid with a first amplification primer containing the target binding sequence SEQ ID NO:1 and a second amplification primer containing the target binding sequence SEQ ID NO:2 or SEQ ID NO:3; and b) the target nucleic acids in the first and second amplification reactions are then elongated on the target nucleic acid. 4. The method of claim 3 also involves the detection of a bridled, amplified target nucleic acid with a detectable probe containing SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12 labeled with a detectable label. 5. Method of claim 3, where the first and second amplified primers also contain recognition regions for restriction estrogen (ES) donucleases that cleave the recognition site. When the recognition site is hemodified and the target nucleic acid is amplified by strand substitution amplified. 6. Method of Claim 3, where the first amplification primer contains the target binding sequence SEQ ID NO:1 and the second amplification primer contains the target binding sequence SEQ ID NO:2 or SEQ ID NO:3, and the target nucleic acid is amplified by polymerase chain reaction. 7. Methods for complex-specific amplification of Mycobacterium avium target nucleic acids include: a) hybridizing a primary amplification primer containing SEQ ID NO:1 and a second amplification primer containing SEQ ID NO:2 or SEQ ID NO:3 with the target nucleic acid; and b) amplifying the target nucleic acid by strand replacement. 8. The method of claim 7 also involves detecting the hybridized target nucleic acid with a detectable probe containing SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12 labeled with a detectable label. 9. Method of claim 7 whereby the amplified target nucleic acid is captured for detection by hybridization with a capture probe containing SEQ ID NO:7 or SEQ ID NO:8 attached to a ligand. 10. Method of Claim 7 where a second amplification primer containing SEQ ID NO:2 is used to replace the first and second amplification primers from the target nucleic acid by the attachment of the first buffer primer containing SEQ ID NO:5 and the second buffer primer containing SEQ ID NO:6 (Claim 10, 2 pages, 0 figures).