Primer and Probe for Use In Detection of Mycobacterium Kansasii and Method for Detection of Mycobacterium Kansasii Using The Same

Abstract

The present invention discloses an oligonucleotide which comprises a part or the entire sequence of the nucleotide sequence depicted in SEQ ID NOS: 1-4, or a part or the entire sequence of a nucleotide sequence complementary to SEQ ID NOS: 1-4, wherein the oligonucleotide is capable of hybridizing with the nucleotide sequence of Mycobacterium kansasii; a primer and a probe for detecting M. kansasii comprising the oligonucleotide; and a method for detecting M. kansasii using the primer and / or probe.The methods enable the detection of M. kansasii more rapidly and with higher accuracy compared with conventional methods performed by culture. Further, the methods can exclude false positive results for the diagnosis and can also detect and diagnose M. kansasii with higher accuracy compared with methods performed by PCR using conventional primers and / or probes. Still further, the method can quantify M. kansasii cells.

Description

RELATED APPLICATIONS[0001]This is a division of application Ser. No. 11 / 920,234, filed Nov. 13, 2007, issued as U.S. Pat. No. 8,242,249, which is a national stage application of PCT Application No. PCT / JP2006 / 309514, filed May 11, 2006, which claims the benefit of JP 2005-141153, filed May 13, 2005, all of which are incorporated herein by reference.TECHNICAL FIELD[0002]The present invention relates to a method for detecting and / or identifying M. kansasii (Mycobacterium kansasii, hereinafter described as M. kansasii) through the use of amplification of nucleic acid and detection system thereof in clinical laboratory test.BACKGROUND ART[0003]Nontuberculous mycobacterium (NTM) is a gram positive bacillus having acid-fast characteristics classified into genus Mycobacterium, and is a sort of acid-fast bacterium other than tuberculosis complex and Mycobacterium leprae. [0004]Among nontuberculous mycobacterium, clinically problematic bacterial strain is known to include Mycobacterium kansa...

AI Technical Summary

Benefits of technology

The method described in this patent allows for faster and more accurate detection of a specific bacteria called M. kansasii compared to traditional methods of identifying bacteria through culture examination. Additionally, this method helps to exclude false positive results and ensures high accuracy in detecting M. kansasii. It can also be used to quantify M. kansasii cells.

Problems solved by technology

However, since nontuberculous mycobacterium disease has no specific clinical symptom, it is quite difficult to differentiate tuberculosis from nontuberculous mycobacterium disease by clinical observation and histopathological manifestation, moreover, to specify the species of the nontuberculous mycobacterium.

By this test, only “positive acid-fast bacterium” can be recognized, and differentiation of tuberculosis bacterium from nontuberculous mycobacterium cannot be achieved.

However, in general, growth of bacterium belonging to genus Mycobacterium is slow, and takes considerable time for its culture.

In addition, there is another problem that it requires additional 2 to 3 weeks to complete various biochemical tests for the identification of bacterial species.

However, since some other species belonging to genus Mycobacterium show the same properties as M. kansasii shows, the identification of M. kansasii by its coloring property is generally of a problem.

In addition, as the judgment is made positive regardless of the size of bacterial count, and since the number of the bacterium is unknown, diagnosis of infectivity whether it is positive or not will be provided with uncertainty.

In addition, since the method has a problem that due to too high sensitivity, the possibility of false positive judgment or the like tends to be made.

2). However, in this study, it has been reported that both pMK1-9 probe and commercially available DNA probe (ACCU-PROBE™) were unable to detect considerable number of strain types of M. kans

Moreover, for example, identification of DNA region effective for detecting M. kansasii has also been studied (for example, Patent Document 1 and the like), however, the present situation is that the method of diagnosis specific to M. kansasii has not been established.

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