JP2001299354A - Nucleic acid for detecting bacterium belonging to genus mycobacterium - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method by which many kinds of bacteria belonging to the genus Mycobacterium can be recognized and detected, and to provide a nucleic acid therefor. SOLUTION: A nucleic acid which is used for detection, can utilize the mutations of the base sequences of regions in the gyrase B genes of many kinds of bacteria belonging to the genus Mycobacterium, and can recognize and detect many kinds of the bacteria belonging to the genus Mycobacterium. For example, a nucleic acid having a base sequence represented by the sequence number 16 as a nucleic acid for detecting Mycobacterium avium.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a nucleic acid for detecting a bacterium belonging to the genus Mycobacterium and a method for detecting a bacterium belonging to the genus Mycobacterium using the nucleic acid.

[0002]

2. Description of the Related Art Conventionally, Mycobacterium genus bacteria such as Mycobacterium tuberculosis have been detected by culturing a sample, growing the bacterium, and examining the mycological properties. However, this method has the disadvantage that it takes time and effort.

[0003] Genetic testing is also generally known as a method for detecting bacteria. However, it is not easy to identify the species of Mycobacterium by genetic testing because of the high homology of the genes of Mycobacterium.

[0004] Recently, it has been proposed to discriminate bacteria by utilizing the gyrase B (gyrB) gene of bacteria (Japanese Patent Application Laid-Open Nos. 11-169175 and 11-290079).
JP-A-11-169175 describes the nucleotide sequence of the region in the gyrB gene of two Mycobacterium genus bacteria, Mycobacterium simiae and Mycobacterium asiaticum. Japanese Patent Application Laid-Open No. H11-290079 discloses that the gyrB gene of four Mycobacterium genus bacteria, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum and Mycobacterium microti. The nucleotide sequence of the region is described, and their differences are also examined.

[0005]

However, there are many other clinically important species of Mycobacterium belonging to the genus Mycobacterium, but the nucleotide sequence of the gyrB gene region is not known at all. Therefore, there is no means for detecting Mycobacterium bacteria other than the above-mentioned six species. Further, there is no known method for species-specific detection of Mycobacterium bacteria, including Mycobacterium bacteria other than the above-mentioned six species.

Accordingly, an object of the present invention is to provide a nucleic acid for identifying and detecting many Mycobacterium genus species and a method for detecting Mycobacterium genus bacteria using the nucleic acid.

[0007]

Means for Solving the Problems As a result of earnest studies, the present inventors have succeeded in determining the nucleotide sequence of the region in the gyrB gene of nine Mycobacterium species other than the above six species. Furthermore, based on such information, a method for species-specific detection of various Mycobacterium bacteria has been conceived, and the present invention has been completed.

[0008] That is, the present invention relates to SEQ ID NO: 1 in the sequence listing.
In the region of 867 nt to 900 nt of the nucleotide sequence represented by, a region of 8 or more consecutive nucleotides, g of 877 nt, a of 879 nt and 88
A nucleic acid for detecting Mycobacterium avium comprising a nucleic acid fragment having a nucleotide sequence of a region containing at least one of 5 nt c; 86 of the nucleotide sequence represented by SEQ ID NO: 2 in the sequence listing;
Mycobacterium intracellulare detection consisting of a nucleic acid fragment consisting of a region consisting of at least one of g of 879 nt, c of 885 nt, and g of 888 nt, which is a continuous region of 8 bases or more in the region of 8 nt to 900 nt A nucleic acid for use; a region of 8 or more consecutive nucleotides in the region of 874 nt to 906 nt of the base sequence represented by SEQ ID NO: 2 in the sequence listing, g of 888 nt, g of 891 nt and
A nucleic acid for detecting Mycobacterium intracellulare consisting of a nucleic acid fragment comprising a region containing at least one of 894 nt g; 86 of the base sequence represented by SEQ ID NO: 3 in the sequence listing
A Mycobacterium comprising a nucleic acid fragment comprising a region of at least 8 contiguous bases in the region of 1 nt to 904 nt and comprising at least one of g of 871 nt, t of 873 nt, t of 879 nt and a of 883 nt. A nucleic acid for detecting Mycobacterium asiaticum, Mycobacterium marinum and / or Mycobacterium sulgai comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 20 in the sequence listing; In the region of SEQ ID NO: 4, 8 or 11, a region of 8 or more consecutive bases in a region extending from 10 bases upstream to 10 bases downstream from the region having the base sequence represented by SEQ ID NO: 20 in the sequence listing Thus, 1 nt t, 3 nt g, 6n in the base sequence represented by SEQ ID NO: 20
Mycobacterium asiaticum, Mycobacterium marinum and / or Mycobacterium comprising a nucleic acid fragment having a nucleotide sequence of a region containing at least one of t a, 9 nt c, 13 nt a and 14 nt c -A nucleic acid for detecting a chimpanzee; a nucleic acid for detecting Mycobacterium asiaticum and / or Mycobacterium gondae composed of a nucleic acid fragment having a base sequence represented by SEQ ID NO: 21 in the sequence listing; SEQ ID NO: 4 or 6 in the sequence listing In the region extending from 10 bases upstream to 10 bases downstream of the region having the base sequence represented by SEQ ID NO: 21 in the sequence listing, a region of 8 or more consecutive bases, A Mycobacterium comprising a nucleic acid fragment having a nucleotide sequence of a region containing at least one of 5 nt t, 8 nt g, 9 nt t and 11 nt g in the nucleotide sequence shown. Jiatikamu and / or Mycobacterium Godonae detector nucleic acid; a continuous 8 bases or more regions in the region of 873nt~906nt the nucleotide sequence represented by SEQ ID NO: 4, c of 885nt, 88
A nucleic acid for detecting Mycobacterium asiaticum comprising a nucleic acid fragment comprising a region containing at least one of 9 nt a and 890 nt c; in the 861 nt to 895 nt region of the base sequence represented by SEQ ID NO: 5 in the sequence listing A contiguous region of 8 bases or more, including 871 nt g, 873 nt t, 876 nt t, and 879 nt
t, a nucleic acid fragment for detecting Mycobacterium gastry consisting of a nucleic acid fragment comprising at least one of a 883 nt a and 884 nt c; a mycobacterium consisting of a nucleic acid fragment having a base sequence represented by SEQ ID NO: 24 in the sequence listing Umm
A nucleic acid for detecting Gordonae and / or Mycobacterium simiae; a region in SEQ ID NO: 6 or 10 in the sequence listing, from the upstream 10 bases to the downstream of the region having the base sequence represented by SEQ ID NO: 24 in the sequence listing A continuous region of 8 or more bases in a region extending over 10 bases,
1 nt t, 3 nt g, 6 nt a, 13 nt in the nucleotide sequence shown by
A nucleic acid for detecting Mycobacterium gordonae and / or Mycobacterium simiae comprising a nucleic acid fragment having a nucleotide sequence of at least one of a and 14nt c; a nucleotide sequence represented by SEQ ID NO: 6 in the sequence listing 873nt ~ 9
A region of 8 or more consecutive nucleotides in the 07 nt region,
A nucleic acid for detecting Mycobacterium gordonae consisting of a nucleic acid fragment comprising at least one of 5 nt t, 889 nt a and 890 nt c; a region of 868 nt to 901 nt of the base sequence represented by SEQ ID NO: 7 in the sequence listing A contiguous region of 8 or more bases, g of 879 nt, t of 882 nt, c of 885 nt, 88
A nucleic acid for Mycobacterium marmoense detection consisting of a nucleic acid fragment comprising a region containing at least one of 9 nt a, 890 nt c and 891 nt t; a region of 872 nt to 907 nt of the base sequence represented by SEQ ID NO: 7 in the sequence listing A nucleic acid for detecting Mycobacterium marmoense comprising a nucleic acid fragment consisting of a region comprising at least any of 885 nt c, 889 nt a, 890 nt c and 891 nt t; 858n of the nucleotide sequence represented by SEQ ID NO: 8 in the sequence listing
a Mycobacterium bacterium comprising a nucleic acid fragment consisting of a region comprising at least any of 873 nt, 876 nt, 877 nt t, and 879 nt g; A nucleic acid for detecting a marinum; a region consisting of 8 or more consecutive nucleotides in the region of 873 nt to 907 nt of the base sequence represented by SEQ ID NO: 8 in the sequence listing, and 885 nt c and 889 nt
a, a nucleic acid fragment for detecting Mycobacterium marinum consisting of a nucleic acid fragment comprising at least one of c of 890 nt and a of 892 nt; in the region of 867 nt to 900 nt of the nucleotide sequence represented by SEQ ID NO: 9 in the sequence listing; Contiguous region of 8 bases or more, t of 877 nt, g of 882 nt, a of 889 nt, and c of 890 nt
A nucleic acid for detecting Mycobacterium scroflaseum, comprising a nucleic acid fragment comprising a region containing at least one of the following: 858 nt to 858 nt of the nucleotide sequence represented by SEQ ID NO: 9 in the sequence listing
It is a continuous region of 8 bases or more in the 91 nt region,
A nucleic acid for detection of Mycobacterium scroflaseum comprising a nucleic acid fragment comprising a region containing at least one of 6 nt a and 877 nt t; contiguous in the 874 nt to 907 nt region of the base sequence represented by SEQ ID NO: 9 in the sequence listing A nucleic acid for detecting Mycobacterium scroflaseum comprising a nucleic acid fragment comprising a region of at least 8 bases and comprising at least one of a 889 nt a to 891 nt g; a nucleic acid having the nucleotide sequence represented by SEQ ID NO: 10 in the sequence listing; A continuous region of 8 bases or more in the region of 873 nt to 907 nt, wherein t is 885 nt, a is 889 nt,
A nucleic acid for detection of Mycobacterium similiae, comprising a nucleic acid fragment comprising a region containing at least one of c of 890 nt and a of 897 nt; contiguous in the region of 859 nt to 891 nt of the base sequence represented by SEQ ID NO: 11 in the sequence listing A nucleic acid for detecting Mycobacterium surugai comprising a nucleic acid fragment comprising a region of at least 8 bases and comprising at least one of g of 876 nt, t of 877 nt, and g of 879 nt; and SEQ ID NO: 11 in the sequence listing. It is a contiguous region of 8 or more bases in the region of 874 nt to 907 nt of the base sequence shown,
A, a 890 nt c and / or a 894 nt c.
Provided is a nucleic acid for detecting chimpanzee.

Further, the present invention relates to SEQ ID NO: 12 in the sequence listing.
A nucleic acid for Mycobacterium tuberculosis detection consisting of a nucleic acid fragment consisting of a contiguous region of 8 or more bases in the 861 nt to 894 nt region of the base sequence; A nucleic acid for detecting Mycobacterium tuberculosis comprising a nucleic acid fragment consisting of a continuous eight or more bases in the region of No. 5; eight or more continuous bases in the region of 817 nt to 851 nt of the base sequence represented by SEQ ID NO: 5 in the sequence listing Area, 831nt g, 834n
a nucleic acid for detection of Mycobacterium gastori comprising a nucleic acid fragment comprising a region containing at least one of c of t and 837 nt of t; a nucleotide sequence represented by SEQ ID NO: 13 in the sequence listing;
A nucleic acid for detecting Mycobacterium bovis consisting of a nucleic acid fragment consisting of a region containing at least 837 nt and being a region of at least 8 consecutive bases in the region of 820 nt to 854 nt; a base represented by SEQ ID NO: 41 in the sequence listing A nucleic acid for detecting Mycobacterium bovis, Mycobacterium microti and / or Mycobacterium africanum comprising a nucleic acid fragment having a sequence; a region in SEQ ID NO: 13, 14 or 15 of the sequence listing, which is a sequence listing From the upstream 10 bases to the downstream 1 to the region having the base sequence represented by SEQ ID NO: 41
A Mycobacterium bovis, a Mycobacterium bovis, a Mycobacterium bovis, comprising a nucleic acid fragment having a base sequence of at least 7 nt in the base sequence represented by SEQ ID NO: 41, which is a continuous region of 8 bases or more in a region extending over 0 bases A nucleic acid for detecting Bacterium microti and / or Mycobacterium africanum; Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium bovis composed of a nucleic acid fragment having a base sequence represented by SEQ ID NO: 42 in the sequence listing. A nucleic acid for detecting Microti and / or Mycobacterium africa; 10 bases upstream of a region having a base sequence represented by SEQ ID NO: 42 in SEQ ID NO: 12, 14 or 15 in the sequence listing Having a nucleotide sequence of a region of 8 or more consecutive nucleotides in a region ranging from 10 nucleotides downstream to A nucleic acid for detecting Mycobacterium tuberculosis, Mycobacterium microti and / or Mycobacterium africanum consisting of an acid fragment; Mycobacterium tuberculosis consisting of a nucleic acid fragment having a base sequence represented by SEQ ID NO: 43 in the sequence listing A nucleic acid for detecting Mycobacterium bovis, Mycobacterium microti and / or Mycobacterium africanum; a region in SEQ ID NO: 12, 13, 14 or 15 of the sequence listing, wherein SEQ ID NO: 43 in the sequence listing A mycobacterium tuberculosis, a mycobacterium bovis consisting of a nucleic acid fragment having a base sequence of a region of 8 or more consecutive bases in a region extending from 10 bases upstream to 10 bases downstream of the region having the base sequence represented by Mycobacterium microti and / or A nucleic acid for detecting Mycobacterium bovis; a nucleic acid for detecting Mycobacterium bovis comprising a nucleic acid fragment consisting of a continuous eight or more nucleotides in the region of 817 nt to 851 nt of the nucleotide sequence represented by SEQ ID NO: 13 in the sequence listing; provide.

[0010] The present invention further provides a method for detecting a bacterium belonging to the genus Mycobacterium, which is performed using a plurality of the nucleic acids for detection of the present invention. Further, the present invention provides a kit for detecting a Mycobacterium bacterium comprising a plurality of the above-described nucleic acids for detection of the present invention.

[0011]

DETAILED DESCRIPTION OF THE INVENTION Mycobacterium avium (M
ycobacterium avium), mycobacterium intracellulare (M. intracellulare), mycobacterium
M. kansasii, M. asiaticum, M. gastri, M. gondae (M. g)
ordonae), Mycobacterium malmoense (M. malm
oense), Mycobacterium marinum (M. marinum),
Mycobacterium scrofulaceum
um), Mycobacterium simiae and M. szulgai, the nucleotide sequences of the regions in the gyrB gene are shown in SEQ ID NOs: 1 to 11, respectively. Also, Mycobacterium tuberculosis
(M.tuberculosis), Mycobacterium bovis
vius), M. african
um) and Mycobacterium microti
The nucleotide sequences of the regions in the gyrB gene of Mycobacterium belonging to the genus Mycobacterium are shown in SEQ ID NOs: 12 to 15, respectively.

By comparing these nucleotide sequences, a fragment containing a site different from the corresponding portion of the sequence of another species can be used for differential detection of Mycobacterium bacteria. When nucleic acids are used for detection of bacteria, a probe or P
Used as a primer in nucleic acid amplification methods such as CR. The probe is provided with an appropriate label such as a fluorescent label, a radiolabel, an enzyme label, and a biotin label. In the present specification, such a labeled nucleic acid is also included in the “nucleic acid for detection”. In addition, a nucleic acid not labeled is carried on a carrier,
The nucleic acid in the sample can also be detected by labeling the nucleic acid in the sample and hybridizing it to the nucleic acid carried on the carrier. In the present specification, the immobilized nucleic acid carried on a carrier without labeling in this way is also included in the “probe”. When a nucleic acid in a sample is detected by labeling, it is preferable to amplify the nucleic acid in the sample by a nucleic acid amplification method in order to increase sensitivity.

When the nucleic acid for detection according to the present invention is used as a primer in a nucleic acid amplification method, the nucleic acid in a sample can be quantified by using so-called real-time PCR or the like as the nucleic acid amplification method. Further, even when used as a probe, quantification or semi-quantification becomes possible using the concentration of the label as an index. As used herein, the term "detection"
Such quantitative or semi-quantitative detection is also included.

Although the size of the nucleic acid for detection of the present invention is not particularly limited, it is usually preferably about 8 to 50 bases. When used as a probe, it is more preferably about 10 to 20 bases.

Particularly preferred specific examples of the nucleic acid for detection of the present invention include 28 kinds of nucleic acids used in the following Examples. These are particularly preferably used as probes. The names, base sequences, sequence numbers of the 28 kinds of probes, the species of Mycobacterium to be detected, the sequence numbers indicating the base sequences of the nucleic acids to which each probe hybridizes, and the site to which each probe hybridizes are as follows. The results are shown in Tables 1 and 2.

[0016]

[Table 1]

[0017]

[Table 2]

Nucleic acid fragments having a base sequence of 10 or more bases upstream to 10 bases downstream of the region to which these probes hybridize, and having a contiguous region of 8 bases or more can also be used as the nucleic acid for detection of the present invention. It can be preferably used. However, it is necessary that the base sequence of each probe shown in Tables 1 and 2 contains at least one base that is different from the base sequence of the corresponding region of another species. Bases that differ from the base sequences of the corresponding regions of other species are indicated by underlining the base sequences in Tables 1 and 2. When there are a plurality of such mutation sites in one probe, it can be used as long as it contains at least one of them.
It is preferable to include all of them because identification can be performed with higher sensitivity.

The nucleic acid for detection of the present invention is preferably used in combination of two or more, thereby enabling identification of Mycobacterium bacteria. In particular, it is preferable to use them in combination so that the number of bacteria to be detected is 7 or more, more preferably 10 or more. In particular, it is preferable to use the probes shown in Tables 1 and 2 in combination. The probes shown in Tables 1 and 2 have been cleverly designed so that a maximum of 14 kinds of Mycobacterium bacteria can be discriminated and easily detected by using them in combination.

The Mycobacterium bacterium detector can be obtained by loading a plurality of the nucleic acids for detection of the present invention on a carrier such as a slide glass, a membrane, or a microtiter plate. In this case, the target gene in the sample is amplified by a nucleic acid amplification method, and labeled simultaneously or after amplification by a conventional method, hybridized with a solid-phased probe, and the label bound to the solid phase is detected. Is preferred. The loading of the probe on the carrier can be performed by a conventional method.

[0021]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below more specifically based on embodiments. However, the present invention is not limited to the following examples.

(1) Determination of the nucleotide sequence of the region in the gyrB gene The Mycobacterium avium (Mycob
acterium avium), Mycobacterium intracellulare, Mycobacterium kansasii, Mycobacterium gastri
(M. gastri), Mycobacterium gordonae (M. gor
donae), M. malmoe (M. malmoe)
nse), Mycobacterium marinum (M. marinum), Mycobacterium scrofulaceum (M. scrofulaceu)
m), and Mycobacterium szulgai (M. szulgai)
The nucleotide sequence of the region in the gyrB gene was determined.

That is, DNA was extracted from the cells of each of the above Mycobacterium species by a conventional method. Using this as a template, a region in the gyrB gene was amplified by PCR using a DNA fragment having the nucleotide sequence of SEQ ID NO: 44 or SEQ ID NO: 45 as a primer in the usual manner, and a commercially available DNA sequencer was used. Thus, the nucleotide sequence of the amplification product was determined.

The determined sequences are represented by SEQ ID NOs: 1-3, 5-9,
11 respectively. For the nucleotide sequence of the region in the gyrB gene of Mycobacterium simiae and Mycobacterium asiaticum, information described in JP-A-11-169175 was used. These sequences are shown in SEQ ID NOs: 4 and 10. In addition, the nucleotide sequence of the region in the gyrB gene of four kinds of Mycobacterium belonging to Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum and Mycobacterium microti is disclosed in The information described in JP 290079 was used. These nucleotide sequences are shown in SEQ ID NOs: 12 to 15.

(2) Preparation of Sample DNA Various Mycobacterium bacteria were cultured by liquid culture in a conventional manner. Thereafter, the cells were collected by centrifugation at 12000 rpm at 4 ° C. for 10 minutes. This was dissolved in 50 ul of purified water and heated at 100 ° C. for 10 minutes. This was centrifuged at 15000 rpm at 4 ° C. for 10 minutes, and the supernatant was transferred to a new tube, which was used as a DNA solution.

(3) PCR Using this DNA as a template, PCR was performed. Primer UP1TL,
The nucleotide sequence of UP2TLr was as follows. 5 'CAYCTCNGGNGGNAARTTYGA 3' 5 'TCNACRTCNGCRTCNGTCAT 3' Reaction composition is 10x PCR buffer (100mM Tris HCl, pH8.
3,500mM KCl, 15mM MgCl _Two, 0.001% (W / V) gelatin） 10
μl, 2mM each dNTP 10μl, 50μM Primer UP1T
L 2μl, 50μM Primer UP2TLr 2μl, AmpliTaq Go
The reaction was performed with 0.5 μl of ld (PERKIN ELMER) and 10 μl of DNA.
The conditions were 96 ° C for 1 minute, 60 ° C for 1 minute, 72 ° C for 2 minutes, and 60 cycles.
I responded. Confirmation of target product by 3% agarose gel electrophoresis
I confirmed.

(4) Solid phase on a slide glass After adjusting each probe shown in Tables 1 and 2 to 200 μM / μl, an equal amount of a spotting solution was added, and a final concentration of 10 μm was added.
It was adjusted to 0 μM / μl. This was dispensed into a microtiter plate and then solid-phased on a slide glass using a microarray maker. After drying overnight, it was washed with 0.2% SDS solution and distilled water. Next, after heating with distilled water of 95 ° C, Na2B
Washed with H4 solution. After washing again with the SDS solution and distilled water, it was dried to prepare a DNA chip.

(5) Labeling of Sample PCR was performed using primers of F-119: TGGGCAACACCGAAGTGAAGTCGTT R-99: CCGAGGACACAGCCTTGTT, and the amplified sample was labeled at that time. After labeling, purification was performed using a Sephadex G-50 column. Reaction composition is 10 × PCR
buffer (100 mM Tris HCl, pH 8.3, 500 mM KCl, 15 mM MgCl
₂ , 0.001% (W / V) gelatin) 10 μl, 10 mM d (ATG) TP & 2.
2 μl of 5 mM dCTP, FluoroLink Cy5-dCTP (1 mM) (trade name, fluorescent substrate manufactured by amershampharmacia biotech) 1 μm
l, 50μM Primer F-119 2μl, Primer R-99
2μl, AmpliTaq Gold (PERKIN ELMER) 0.6μl, DNA 5μ
1 Conditions are 96 ° C for 1 minute, 60 ° C for 1 minute, 72 ° C for 4 minutes.
The reaction was performed for 60 minutes per minute. After PCR labeling, the labeled product was purified using a Sephadex G-50 column.

(6) Hybridization A purified Cy-5 label was added to the DNA chip prepared above,
The reaction was performed for 4.18 hours. 2 × SSC-0.1% SDS washing solution was added, and washed three times at room temperature for 10 minutes. Next, 0.1 × SSC-0.1%
After washing three times with the SDS washing solution, the plate was washed with 0.1 × SSC at 50 ° C. for 10 minutes. Let dry. This was scanned with a microarray reader and the data was analyzed.

(7) Results The results are shown in the following Tables 3 to 5 as analysis tables. A circle indicates that the sample nucleic acid hybridized with the immobilized probe.

[0031]

[Table 3]

[0032]

[Table 4]

[0033]

[Table 5]

[0034]

As described above, according to the present invention, discriminative detection of Mycobacterium bacteria, which has been difficult so far, can be performed easily and accurately.

[0035]

[Sequence List] SEQUENCE LISTING <110> SRL, INC. <120> Nucleic Acids for Detecting Bacteria Belonging to Genus Mycobacterium <130> 00652 <160> 47

[0036] <210> 1 <211> 1263 <212> DNA <213> Mycobacterium avium <400> 1 ggcgagaaca gcggctacaa cgtcagcggc ggtctgcacg gcgtcggcgt ctcggtggtc 60 aacgcgctgt ccactcggct cgaggtcaac atcgcccgcg acggctacga gtggtcgcag 120 tactacgacc acgccgtgcc cggcaccctc aagcagggcg aggccaccaa gcgcaccggc 180 accaccatcc ggttctgggc cgaccccgac atcttcgaga ccaccgagta cgacttcgaa 240 acggtggccc ggcggctgca ggaaatggcg ttcctcaaca agggcctgac catcaacctc 300 accgacgagc gggtgaccaa cgaagaggtc gtcgacgagg tggtcagcga caccgccgac 360 gcacccaagt cggcgcagga gaaggcggcg gaatcggctg cgccgcataa ggtcaagcac 420 cgcaccttcc actaccccgg cggcctggtc gacttcgtca aacacatcaa tcgcaccaaa 480 aaccccatcc accagagcat catcgatttc ggtgggaagg gccccggcca cgaggtcgag 540 atcgcgatgc agtggaacgg cggctactcc gaatcggtgc acaccttcgc caacaccatc 600 aacacgcacg agggcggcac ccacgaggag ggcttccgca gcgcgctgac ctccgtggtc 660 aacaagtacg ccaaggacaa gaagctgctc aaggacaagg accccaacct gaccggcgac 720 gacatccgcg agggtttggc cgcggtgatc tcggtcaagg tgagcgaacc gcagttcgag 780 ggccagacc a agaccaaact gggcaacacc gaggtgaagt cgttcgtgca gaaggtgtgc 840 aacgaacagc tcacccactg gttcgaagcc aaccccgcag acgccaaagt cattgtcaac 900 aaggcggttt catcagcgca ggcgcgcatc gccgcgcgca aggcgcgaga gttggtgcgc 960 cgcaagagcg caaccgacct gggcgggctg cccggcaagc tcgccgactg ccggtcgacc 1020 gacccgcgca agtcggaatt gtatgtggtc gagggtgact cggccggcgg ctcggcgaaa 1080 agcggccggg actcgatgtt ccaggccatc cttccgctgc gcggcaagat catcaacgtc 1140 gaaaaggccc gcatcgaccg ggtgctgaag aacaccgagg tgcaggcgat catcaccgcg 1200 ctgggcaccg ggattcacga cgagttcgac atcaccaagc tgcgctacca caagatcgtg 1260 ttg 1263

[0037] <210> 2 <211> 1263 <212> DNA <213> Mycobacterium intracellulae <400> 2 ggtgagaaca gcggttacaa cgtcagcggt ggcctgcacg gcgtgggcgt ctcggtggtc 60 aacgcgctgt cgacccggct cgaggtggac atcgcccgcg atggctacga atggtcgcag 120 ttctacgacc acgccgtacc cggaacgctc aaacagggtg aggccaccaa gcggacgggc 180 accacgatca ggttctgggc cgaccccgac atcttcgaga ccaccgagta cgacttcgag 240 acggtggcgc gccggctgca ggaaatggcg ttcctcaaca aggggttgac catcaacctc 300 accgacgagc gggtgagcaa cgaggaggtc gtcgacgagg tcgtcagcga taccgccgac 360 gcacccaagt cggcccagga aaaggcggcg gaatcgactg cgccacataa ggttaagcac 420 cgcaccttcc actaccccgg cggtctggtc gacttcgtca agcacatcaa ccgcaccaag 480 agcccgatcc agcagagcat catcgacttc gacggcaaag gtcccggcca cgaggtcgag 540 atcgcgatgc agtggaacgg cggctactcg gaatccgtgc acaccttcgc caacaccatc 600 aacacccacg agggcggcac ccacgaagag ggcttccgca gcgcgctgac gtcggtggtg 660 aacaagtacg ccaaagacaa gaagttgctg aaagacaagg acccgaacct caccggcgac 720 gacattcgcg aaggcctggc cgcggtgatc tcggtcaagg tcagcgaacc gcagttcgag 780 g gtcagacca agaccaagct gggcaacacc gaagtgaagt cgttcgtgca gaaggtctgc 840 aacgaacagc tcacccactg gttcgaggcc aaccccgcgg acgccaaggt ggtggtcaac 900 aaggcggtgt cgtcggcgca ggcccggatc gccgcgcgca aggcgcgaga gttggtgcgt 960 cgcaagagcg ccaccgatct gggcgggctg cccggcaagc tcgccgactg ccgctcgacg 1020 gatccgcgca agtcggaact gtatgtggtg gagggtgatt cggccggcgg ctcggcgaag 1080 agcggccgcg actcgatgtt ccaggccatc ctgccgctgc gcggcaagat catcaacgtc 1140 gagaaggccc gcatcgaccg ggtgttgaag aacaccgagg tgcaggccat catcaccgcc 1200 ctgggcaccg gcatccacga cgagttcgac atcaccaagc tgcgctatca caagatcgtg 1260 ctg 1263

[0038] <210> 3 <211> 1257 <212> DNA <213> Mycobacterium kansasii <400> 3 tccgacgcct acgcgatatc gggcgggctg cacggtgtgg gtgtctcggt ggtcaacgca 60 ctgtccaccc ggctggaggt ggagatcaag cgcgacggcc atgagtggtc gcaggtttac 120 gagaaatccg agccgatggg actcaagcaa ggcgcgccga ctaagaagac cggcacgacg 180 gtgcggttct gggccgatcc caatgttttt gagaccaccg agtacgactt cgaaaccgtc 240 gcacgacggt tgcaggagat ggcgtttctc aacaaggggc tcaccatcaa tctgaccgat 300 cagcgggtga cccaggacga ggtcgtcgac gaggtggtca gcgacgtcgc cgaggcccca 360 aagtcggcca gcgagaaggc ggccgaatcc gccgccccgc acaaggtcaa gaagcgtacc 420 ttccactatc ccgggggtct ggttgacttc gtcaagcaca tcaaccggac caagaacgcc 480 atccacagca gcatcgtcga cttctccggt aagggacccg gccacgaagt ggagatcgcg 540 atgcagtgga atgccggcta ttcggagtcg gtgcatacct tcgccaacac catcaacacc 600 cacgagggtg ggacccacga agaggggttc cgcagcgcgc tgacctcggt ggtgaacaag 660 tacgccaagg accgcaaact gctcaaggaa aaggacccca acctcaccgg cgacgacatc 720 cgggaagggt tggccgcggt gatttcggtc aaggtcagcg agccgcagtt cgagggccag 780 accaag acga aactgggcaa caccgaggtg aagtcgttcg tgcagaaggt gtgcaacgaa 840 cagctcaccc attggttcga ggccaacccc gctgacgcta aaaccgttgt caacaaggcg 900 gtttcatcgg cgcaagcacg cattgcggcc cgcaaggcgc gcgagttggt gcgccgcaag 960 agcgcaaccg atctgggcgg actacccggc aagctcgccg actgccgctc gaccgacccg 1020 cgcaagtccg aactgtatgt ggtggagggt gattcagccg gcggctcggc gaagagcggt 1080 cgcgactcga tgttccaggc catcttgccg ttgcgcggca agatcatcaa cgtcgagaag 1140 gcccgcatcg accgggtgct gaagaacacc gaagtccagg cgatcatcac cgcgttgggt 1200 accggcatcc acgacgaatt cgacatcgcg agactgcgtt accacaagat cgtgctc 1257

[0039] <210> 4 <211> 1263 <212> DNA <213> Mycobacterium asiaticum <400> 4 ggtgagaaca gcggctacac cgtcagcggt gggctgcacg gtgtcggtgt gtcagtggtc 60 aacgcgttgt cgacccgact cgaggtcgac atcaagcgcg acgggcacga gtggtcccag 120 tattacgagc gcgccgttcc tggcacgctc aagcagggcg aggcgaccaa gaagaccggc 180 accaccatcc ggttctgggc ggacccggac atcttcgaga ccacccagta cgacttcgag 240 acggtggcgc gccggctcca agagatggcg ttcctgaaca agggcttgac catcaacttg 300 accgacgagc gggtggacca ggacgaggtc gtcgatgaag tcgtcagcga caccgccgat 360 gcgcccaagt ccgccgaaga gaaggcggcc gaatccaaag cgccgcacaa ggttaagcac 420 cgcaccttcc actaccccgg cggcttggtc gacttcgtca agcacatcaa ccggaccaag 480 agcccgatcc aacagagcgt catcgacttc gagggcaaag gcaccggcca cgaggtcgag 540 atcgcgatgc agtggaacgg tggctactcg gagtcggtgc acaccttcgc caacacgatc 600 aacacccacg agggcggtac gcacgaagaa gggttccgca gtgcgctgac gtcggtggtg 660 aacaaatacg ccaaagacaa gaagctgctg aaagacaagg acccgaacct caccggtgac 720 gacatccgcg agggactggc cgcggtgatc tcggtcaagg tcgccgaacc ccagttcgag 780 ggcca gacaa agaccaagct gggcaacacc gaggtcaagt cgttcgtgca gaaggtgtgc 840 aacgaacagc tcacccactg gttcgaggcc aatccgtcgg aagccaaaac cgttgtcaac 900 aaggcggttt cgtccgcaca ggcccggatc gcggcgcgga aggcccgaga gttggtgcgg 960 cgcaagagcg cgaccgattt gggcgggctg cccggcaagc tggccgactg ccgttccacc 1020 gacccgcgca agtccgaact gtatgtggtg gagggtgact cggcaggtgg ctcggccaag 1080 agcggccgtg actcgatgtt ccaggccatc ctgccgctgc gcggcaagat catcaacgtc 1140 gagaaggccc gcatcgaccg ggtcctgaag aacaccgaag tccaggcgat catcaccgcg 1200 ctgggtaccg gtattcacga cgagttcgac atttctaaac tgcgttacca caagatcgtg 1260 ttg 1263

[0040] <210> 5 <211> 1257 <212> DNA <213> Mycobacterium gastri <400> 5 tccgacgcct atgcgatatc gggtggactg cacggtgtgg gtgtctcggt ggtcaacgcg 60 ctgtccatcc ggctggaggt ggagatcaag cgcgacggcc atgagtggtc gcaagtttat 120 gagaagtccg agccgatggg actcaagcaa ggcgcgccga cgaagaagac cggcacgacg 180 gtgcggttct gggccgaccc caacgttttt gaaaccaccg agtacgactt cgaaaccgtc 240 gcgcgacggt tgcaggagat ggcgtttctc aacaaggggc tcaccatcaa cctgaccgat 300 cagcgggtaa cccaggacga agtggtcgac gaggtggtca gcgacgtcgc cgaggccccg 360 aagtcggcca gtgagaaggc ggccgaattc accgcccccc acaaggtgaa gaagcgtacc 420 tttcactatc ccggtggctt ggttgacttc gtcaagcaca tcaaccgcac caagaacgcc 480 atccacagca gcatcgtcga cttctccgga aaggggaccg gccacgaagt ggagatcgcg 540 atgcagtgga atgccggcta ttcggagtcg gtgcacacct tcgccaacac catcaacacc 600 catgagggcg ggacccatga agaagggttc cgcagcgcgc tcacgtccgt ggtgaacaag 660 tacgccaagg accgcaaact gctcaaagac aaggacccca acctcaccgg cgacgacatc 720 cgggaagggt tggccgcggt gatttcggtc aaagtcagcg aaccgcagtt cgagggccag 780 accaagac ga aactaggcaa caccgaggtg aagtcgttcg tgcagaaggt gtgcaatgaa 840 cagctcaccc attggttcga ggccaacccc gctgatgcta aaaccgttgt caacaaggca 900 gtttcatcgg cgcaggccag gattgcggcc cgcaaggcgc gcgagttggt gcgccgcaag 960 agcgcaaccg atctgggcgg actaccgggc aagttggccg actgccgctc gaccgacccc 1020 cgtaagtccg aattatatgt ggtggagggt gattcagccg gcggctcggc gaagagcggc 1080 cgcgactcga tgtttcaagc gatcttgccg ttgcgcggca agatcatcaa cgtcgagaag 1140 gcccgcatcg accgggtgct gaagaacacc gaagtccagg cgatcatcac cgcgttgggc 1200 accggtattc acgacgaatt cgacatcgcg agactgcgtt accacaagat cgtgctg 1257

[0041] <210> 6 <211> 1263 <212> DNA <213> Mycobacterium gordonae <400> 6 ggcgagaaca gcggctacac ggtcagcggt gggttgcacg gcgtgggcgt gtcggtggtt 60 aacgcgttgt cgacgcggtt ggaagtcgac atcaaacgcg acgggcacga gtggtcgcag 120 tattacaagc gcgcggtgcc gggcaccctc aagcagggtg agacgacccg caagaccggc 180 accacaatcc ggttctgggc ggatccggag atcttcgaga ccacccaata cgacttcgag 240 acggtggcgc gccggctgca ggagatggcg ttcctgaaca agggtctgac gatcaatctg 300 accgacgaac gcgtcgagca ggacgaggtt gtcgacgagg tcgtcagcga caccgccgaa 360 gcgcccaaat ccgccgaaga gaaggctgcc gaatccaagg ccccgcacaa ggtcaagcag 420 cgcaccttcc actatcccgg tggtctggtc gacttcgtca aacacatcaa ccgcaccaaa 480 agcccgatcc agcagagcgt catcgacttc gaaggcaaag gcaccggcca cgaggtcgaa 540 atcgcgatgc agtggaacgg cggctactcc gaatcggtgc acaccttcgc caacaccatc 600 aacacccacg agggcggcac ccacgaagag ggcttccgca gtgcgctgac ctcggtggtc 660 aacaagtacg ccaaagacaa gaagctgctc aaggagaagg acccgaatct caccggtgac 720 gacatccggg aggggttggc cgcggtgatc tcggtgaagg tcgccgaacc gcagttcgag 780 ggtcag acca agaccaagct gggcaacacc gaggtcaagt cgttcgtgca gaaggtgtgc 840 aacgaacagc tcacccactg gttcgaggcc aatccgtcgg aagctaaaac cgttgtgaac 900 aaagcggtgt cgtccgccca ggcgcggatc gccgcgcgca aagcgcgaga gctggtgcgc 960 cgcaagagcg caaccgacct cggcggcctg ccgggcaagc tcgccgactg ccgttcgacg 1020 gatccccgca aatccgaact gtatgtggtg gagggggact ccgccggcgg ctcggccaag 1080 agcggtcggg attcgatgtt ccaggcgatt cttccgttgc gcggcaagat catcaacgtc 1140 gagaaggccc gcatcgaccg ggtgctgaag aacaccgaag tccaggccat catcaccgcg 1200 ctgggcaccg ggatccacga cgagttcgac atcaccaaac tgcgctacca caagatcgta 1260 ttg 1263

[0042] <210> 7 <211> 1263 <212> DNA <213> Mycobacterium malmoense <400> 7 ggcgagaaca gcggatacaa cgtcagtggc ggtttgcacg gtgtcggcgt gtcggtggtc 60 aacgcgttgt cgacccggct cgaggtggat gtcgcccgcg acggctacat gtggtcacag 120 ttctacgatc acgccgagcc gggaaccctc aaacagggcg aggccaccaa gacgacggga 180 accaccatca ggttctgggc cgatcccgac atcttcgaga ccaccgagta cgacttcgag 240 acggtggcgc gccgactgca ggaaatggcg ttcctgaaca agggtttgac gatcaacctc 300 accgacgagc gggtcagtga agaggaggtc gtcgacgatg tcgtcagcga caccgccgag 360 gcacccaagt ccgccgtaga aaaagcggcc gaatcgactg gcccacacaa ggttaagcac 420 cgcacgttcc actacccggg cggcttggtg gacttcgtca agcacatcaa tcggaccaag 480 aacccgattc acaacagcat cgtggatttc tccggcaagg gaccgggcca cgaggtcgaa 540 atcgcgatgc agtggaatgc cggctactcg gagtcggtgc acaccttcgc caacaccatc 600 aacacccacg agggcggcac ccacgaagag ggcttccgca gcgcgttgac gtcggtggtc 660 aacaaatacg ccaaggaccg caaactcctg aaggacaaag accccaacct caccggcgac 720 gacatccggg aaggcctggc agcggtcatt tccgtcaagg tcagcgaacc gcaattcgag 780 ggcca gacca aaaccaagct gggcaacacc gaggtcaagt cgttcgtgca gaaggtctgc 840 aacgaacagc tcacgcactg gttcgaagcc aacccggcgg atgccaaaac tgttgtaaac 900 aaggcggttt cgtcggccca ggcccgaatc gcagcgcgca aggcgcgaga actggtgcgc 960 cgcaagagcg ccaccgacct cggtgggctg ccgggtaagc tcgcagactg ccgctccacc 1020 gacccgcgaa agtcggaact gtatgtggtg gagggtgact cggccggcgg ctcggccaag 1080 agcggccgcg actcgatgtt ccaggcgatc ctcccgctgc gtggcaagat catcaacgtc 1140 gagaaggcgc gcatcgaccg ggtgctgaag aacaccgaag ttcaggcgat catcaccgcg 1200 ctgggcacgg ggattcacga cgagttcgac atcaccaagc tccggtacca caagatcgtg 1260 ctg 1263

[0043] <210> 8 <211> 1263 <212> DNA <213> Mycobacterium marinum <400> 8 ggcgagaaca gtggttacaa cgtcagtggt ggtctgcacg gcgtgggtgt gtcggtggtc 60 aacgcgctgt ccacccgact ggaagtcgac atcaagcgcg acggatacga gtggtcgcag 120 ttctacgacc gcgcccagcc gggcaccctc aaacagggcg aggcaaccaa gaagaccgga 180 accaccatcc ggttctgggc cgactcggac atctttgaga ccaccgaata cgacttcgag 240 acggtggcgc ggcgcctgca ggagatggcg ttcctcaaca agggcctgac catcaacctc 300 accgacgagc gggtcacccc ggacgaggtc gtcgacgacg tcgtcagtga taccgccgaa 360 gcaccaaagt ccgcccagga gaaggccgcc gaatcgaccg cgccgcacaa ggtcaagagc 420 cgcaccttcc actatcccgg cggtttggtc gatttcgtca agcacatcaa ccgcaccaag 480 agtccgattc agcagagcat cgtcgacttc gagggcaagg gctccggcca cgaagtcgaa 540 atcgcgatgc agtggaacgg cggctactcg gagtcggtgc acaccttcgc caacaccatc 600 aacacccatg agggtggaac gcacgaagag ggcttccgca gtgcgttgac ctcggtggtg 660 aacaagtacg ccaaagacaa gaagctgctc aaggacaagg accccaacct caccggtgac 720 gacatccgcg aggggttggc cgcggtcatc tcggtgcggg tggcagagcc gcagttcgag 780 ggtcaga cga agaccaagct gggcaacacc gaggtcaagt cgtttgtcca gaaggtttgt 840 aacgagcagc tcacccactg gttcgaggcc aatccttcgg aagccaaaac cattgtgaac 900 aaggcggtat cctcggcgca ggcacgtctc gccgcgcgca aggcgcgaga gttggtgcgt 960 cgcaagagcg caaccgatct cggtgggctg cccggcaagt tggccgactg ccgctcgaca 1020 gatccgcgta agtcggaact gtatgtggtg gagggtgact cggccggcgg ctcggcaaag 1080 agtggccgcg attcgatgtt ccaggcgatc ctgccgctgc gcggcaagat catcaatgtc 1140 gaaaaggcac gcatcgaccg agtcctgaaa aacactgaag tccaggcgat catcaccgcg 1200 ttgggtaccg gtattcacga cgaattcgac ctctcgaagc tgcgctatca caagatcgtc 1260 ttg 1263

[0044] <210> 9 <211> 1263 <212> DNA <213> Mycobacterium marinum <400> 9 ggcgagaaca gcggctacac cgtcagcggt gggttgcacg gagtgggcgt gtcggtggtc 60 aacgcgctgt ccacccgcct ggaggtcacc atcaagcgcg acgggcacga gtggtttcag 120 tactacgacc gcgccgtgcc cggaaccctc aagcagggcg aggccaccaa gaagaccgga 180 accacgatca ggttctgggc ggaccccgaa atcttcgaaa ccacacagta cgacttcgag 240 accgtggcgc ggcggctgca ggagatggcc ttcctcaaca agggcctcac catcaacctc 300 accgacgaac gagtggagca ggacgaggtc gtcgacgagg tcgtcagcga caccgccgag 360 gcaccgaagt ccgccgaaga gaaggccgcg gaatcgactg cgccacacaa ggtcaagcac 420 cgcaccttcc actaccccgg cggtctggtc gacttcgtca agcacatcaa ccgcaccaag 480 agcccgatcc agcagagcgt catcgatttc gacggcaagg gcaccggcca cgaggtcgag 540 atcgccatgc agtggaacgg cggctactcg gagtccgtcc acaccttcgc caacaccatc 600 aacacgcacg agggcggcac ccacgaggag ggcttccgca gcgcgctgac gtcggtggtg 660 aacaagtacg ccaaagacaa gaaactgctg aaggacaaag atcccaacct caccggtgac 720 gacatccgtg agggcttggc cgcggtcatc tcggtgaagg tcgccgagcc acagttcgaa 780 ggccaga cca agacaaagct gggcaacacc gaggtgaagt cgttcgtgca gaaggtgtgc 840 aacgagcagc tcacccactg gttcgaggcc aacccatccg aggcgaaaac ggtggtgaac 900 aaagcggtgt cgtcggctca ggcgcgcatt gccgcccgca aggcgcgtga actggtgcgc 960 cgcaagagcg ccaccgacct cggcggtctg cccgggaagc tggccgactg ccgctccacc 1020 gacccgcgga aatcggaact gtatgtggtg gagggcgatt cggccggcgg ctcggccaag 1080 agcgggcgcg actcgatgtt ccaggcgatc ctgccgctgc gcggcaagat catcaatgtc 1140 gagaaggccc gcatcgaccg ggtgctgaag aacaccgaag ttcaggcgat catcaccgcg 1200 ctgggtaccg ggattcacga cgagttcgac atcaccaagc tgcgctatca caagatcgtg 1260 ctg 1263

[0045] <210> 10 <211> 1263 <212> DNA <213> Mycobacterium simiae <400> 10 ggggagaaca gtggctacac cgtcagcggc gggttgcacg gggtcggagt gtcggtggtc 60 aacgccctgt ccacccgcct ggaagtcaac gtcaagcgtg acggctatga gtggttccag 120 tactacgacc gggcggtgcc cggcaccctc aagcaaggcg aggcgaccaa gaagaccggc 180 accacgatcc ggttctgggc cgatcctgag atcttcgaaa ccacccagta cgacttcgag 240 acggtggcgc gccggttgca ggaaatggcg ttcctcaaca agggcctgac catcaacctc 300 accgacgaac gtgtcgagca ggacgaggtg gtcgatgagg tggttagcga caccgccgag 360 gcgccgaagt cagccgagga gcaggcggcc gaatcggcca agccgcacaa ggtcaagcac 420 cgcacgttcc actacccggg tgggttggtg gatttcgtca agcacatcaa tcgcaccaaa 480 aacccgatcc agcagagcgt catcgacttc gacggcaaag gaaccgggca cgaagtcgag 540 atcgcgatgc agtggaacgg tggttattcg gagtcggtgc acaccttcgc caacaccatc 600 aacacccatg agggcggcac ccacgaggag ggcttccgca gcgcgctgac ctcggtggtg 660 aacaagtacg ccaaagacaa gaagctgctc aaggacaagg atcccaacct caccggcgac 720 gacatccgag aagggctggc cgcggtgatc tccgtgaagg tcgccgagcc gcagttcgag 780 ggccag acta agacgaaact cggcaacacc gaggtcaagt cgtttgtcca gaaagtctgt 840 aacgaacaac tcactcactg gttcgaggcg aacccgtcgg aagctaaaac cgttgtaaac 900 aaggcggttt cgtcggccca ggcccgcatt gcggcgcgta aggcgcggga gttggtgcgg 960 cgtaagagtg ctacggattt gggtgggttg ccgggcaagt tggctgattg ccgctcgacg 1020 gatccgcgga agtctgagct gtatgtggtg gaaggtgatt ccgcgggtgg gtcggcgaaa 1080 agtgggcgtg attcgatgtt ccaggcgatc ttgccgctgc gcggcaagat catcaacgtc 1140 gaaaaggccc gcatcgatcg ggtgctgaaa aacaccgaag tccaggccat catcaccgcg 1200 ctgggcaccg gcatccacga cgaattcgac atcaccaaac tgcgttacca caagatcgtg 1260 ttg 1263

[0046] <210> 11 <211> 1263 <212> DNA <213> Mycobacterium szulgai <400> 11 ggcgagaaca gtggctacaa cgtcagtggt ggtctgcacg gcgtcggggt gtcggtggtg 60 aacgcgctgt cgacccggct cgaggtcgac atcaagcgtg acggccacaa gtggtcgcag 120 ttctacaaca aggccgtgcc gggcacgctc aaacagggtg aagccactaa gaaaaccgga 180 acgacaatta ggttctgggc cgacccggac atcttcgaga ccaccgaata cgacttcgag 240 accgtggcac gccggctgca ggaaatggca ttcctgaaca agggcttgac catcaacctc 300 accgacgagc gagttgccca ggacgaggtt gtcgacgagg tcgtcagcga caccgccgag 360 gcacccaagt ccgccgaaga aaaggcggcc gaatccaaag ggccgcataa ggttaagcac 420 cgcactttcc attaccccgg cgggctgatc gacttcgtca agcacatcaa ccggaccaag 480 agcccgatcc agcagagtgt cgtcgccttc gacggcaagg gtgaagggca cgaggtcgag 540 atcgcgatgc agtggaacgg cggctattcg gagtcggtgc acaccttcgc caacaccatc 600 aacacccacg agggcggcac ccacgaagaa gggttccgca gcgcactgac atcggtggtg 660 aacaagtacg ccaaagacaa gaagctgctc aaggagaagg acgccaacct caccggcgac 720 gacattcgcg agggcctggc cgcggtcatc tcggtgaaag ttgccgaacc gcagttcgag 780 ggcca gacca agaccaaact gggtaacacc gaggtcaagt cgttcgtaca gaaggtctgc 840 aacgaacagc tgacccactg gttcgaggcc aacccgtcgg aagccaaaac cgtcgtgaac 900 aaggcggtct cgtcggcaca ggcgcgtatc gccgcccgca aggcacgaga gttggtgcgt 960 cgcaagagcg ctaccgatct cggtgggctg cccggcaagc tggccgactg ccgctccacc 1020 gatccgcgca agtcggaatt gtatgtggtg gaaggggact cggccggcgg ctccgccaag 1080 agcggccgcg actcgatgtt tcaggcgata cttccgttgc gcggcaagat catcaacgtc 1140 gagaaggccc gcatcgaccg ggtgctgaag aacaccgaag tccaggcgat catcaccgcg 1200 ctgggtaccg gaattcacga cgagttcgac ctcgccaaac tgcgctacca caagatcgtg 1260 ctg 1263

[0047] <210> 12 <211> 1257 <212> DNA <213> Mycobacterium tuberculosis <400> 12 tcggacgcgt atgcgatatc tggtggtctg cacggcgtcg gcgtgtcggt ggttaacgcg 60 ctatccaccc ggctcgaagt cgagatcaag cgcgacgggt acgagtggtc tcaggtttat 120 gagaagtcgg aacccctggg cctcaagcaa ggggcgccga ccaagaagac ggggtcaacg 180 gtgcggttct gggccgaccc cgctgttttc gaaaccacgg aatacgactt cgaaaccgtc 240 gcccgccggc tgcaagagat ggcgttcctc aacaaggggc tgaccatcaa cctgaccgac 300 gagagggtga cccaagacga ggtcgtcgac gaagtggtca gcgacgtcgc cgaggcgccg 360 aagtcggcaa gtgaacgcgc agccgaatcc actgcaccgc acaaagttaa gagccgcacc 420 tttcactatc cgggtggcct ggtggacttc gtgaaacaca tcaaccgcac caagaacgcg 480 attcatagca gcatcgtgga cttttccggc aagggcaccg ggcacgaggt ggagatcgcg 540 atgcaatgga acgccgggta ttcggagtcg gtgcacacct tcgccaacac catcaacacc 600 cacgagggcg gcacccacga agagggcttc cgcagcgcgc tgacgtcggt ggtgaacaag 660 tacgccaagg accgcaagct actgaaggac aaggacccca acctcaccgg tgacgatatc 720 cgggaaggcc tggccgctgt gatctcggtg aaggtcagcg aaccgcagtt cgagggccag 780 accaagacca agttgggcaa caccgaggtc aaatcgtttg tgcagaaggt ctgtaacgaa 840 cagctgaccc actggtttga agccaacccc accgacgcga aagtcgttgt gaacaaggct 900 gtgtcctcgg cgcaagcccg tatcgcggca cgtaaggcac gagagttggt gcggcgtaag 960 agcgccaccg acatcggtgg attgcccggc aagctggccg attgccgttc cacggatccg 1020 cgcaagtccg aactgtatgt cgtagaaggt gactcggccg gcggttctgc aaaaagcggt 1080 cgcgattcga tgttccaggc gatacttccg ctgcgcggca agatcatcaa tgtggagaaa 1140 gcgcgcatcg accgggtgct aaagaacacc gaagttcagg cgatcatcac ggcgctgggc 1200 accgggatcc acgacgagtt cgatatcggc aagctgcgct accacaagat cgtgctg 1257

[0048] <210> 13 <211> 1257 <212> DNA <213> Mycobacterium bovis <400> 13 tcggacgcgt atgcgatatc tggtggtctg cacggcgtcg gcgtgtcggt ggttaacgcg 60 ctatccaccc ggctcgaagt cgagatcaag cgcgacgggt acgagtggtc tcaggtttat 120 gagaagtcgg aacccctggg cctcaagcaa ggggcgccga ccaagaagac ggggtcaacg 180 gtacggttct gggccgaccc cgctgttttc gaaaccacgg aatacgactt cgaaaccgtc 240 gcccgccggc tgcaagagat ggcgttcctc aacaaggggc tgaccatcaa cctgaccgac 300 gagagggtga cccaagacga ggtcgtcgac gaagtggtca gcgacgtcgc cgaggcgccg 360 aagtcggcaa gtgaacgcgc agccgaatcc actgcaccgc acaaagttaa gagccgcacc 420 tttcactatc cgggtggcct ggtggacttc gtgaaacaca tcaaccgcac caagaacgcg 480 attcatagca gcatcgtgga cttttccggc aagggcaccg ggcacgaggt ggagatcgcg 540 atgcaatgga acgccgggta ttcggagtcg gtgcacacct tcgccaacac catcaacacc 600 cacgagggcg gcacccacga agagggcttc cgcagcgcgc tgacgtcggt ggtgaacaag 660 tacgccaagg accgcaagct actgaaggac aaggacccca acctcaccgg tgacgatatc 720 cgggaaggcc tggccgctgt gatctcggtg aaggtcagcg aaccgcagtt cgagggccag 780 accaaga cca agttgggcaa caccgaggtc aaatcgtttg tgcagaaggt ctgtaatgaa 840 cagctgaccc actggtttga agccaacccc accgactcga aagtcgttgt gaacaaggct 900 gtgtcctcgg cgcaagcccg tatcgcggca cgtaaggcac gagagttggt gcggcgtaag 960 agcgccaccg acatcggtgg attgcccggc aagctggccg attgccgttc cacggatccg 1020 cgcaagtccg aactgtatgt cgtagaaggt gactcggccg gcggttctgc aaaaagcggt 1080 cgcgattcga tgttccaggc gatacttccg ctgcgcggca agatcatcaa tgtggagaaa 1140 gcgcgcatcg accgggtgct aaagaacacc gaagttcagg cgatcatcac ggcgctgggc 1200 accgggatcc acgacgagtt cgatatcggc aagctgcgct accacaagat cgtgctg 1257

[0049] <210> 14 <211> 1257 <212> DNA <213> Mycobacterium microti <400> 14 tcggacgcgt atgcgatatc tggtggtctg cacggcgtcg gcgtgtcggt ggttaacgcg 60 ctatccaccc ggctcgaagt cgagatcaag cgcgacgggt atgagtggtc tcaggtttat 120 gagaagtcgg aacccctggg cctcaagcaa ggggcgccga ccaagaagac ggggtcaacg 180 gtgcggttct gggccgaccc cgctgttttc gaaaccacgg aatacgactt cgaaaccgtc 240 gcccgccggc tgcaagagat ggcgttcctc aacaaggggc tgaccatcaa cctgaccgac 300 gagagggtga cccaagacga ggtcgtcgac gaagtggtca gcgacgtcgc cgaggcgccg 360 aagtcggcaa gtgaacgcgc agccgaatcc actgcaccgc acaaagttaa gagccgcacc 420 tttcactatc cgggtggcct ggtggacttc gtgaaacaca tcaaccgcac caagaacgcg 480 attcatagca gcatcgtgga cttttccggc aagggcaccg ggcacgaggt ggagatcgcg 540 atgcaatgga acgccgggta ttcggagtcg gtgcacacct tcgccaacac catcaacacc 600 cacgagggcg gcacccacga agagggcttc cgcagcgcgc tgacgtcggt ggtgaacaag 660 tacgccaagg accgcaagct actgaaggac aaggacccca acctcaccgg tgacgatatc 720 cgggaaggcc tggccgctgt gatctcggtg aaggtcagcg aaccgcagtt cgagggccag 780 accaa gacca agttgggcaa caccgaggtc aaatcgtttg tgcagaaggt ctgtaacgaa 840 cagctgaccc actggtttga agccaacccc accgactcga aagtcgttgt gaacaaggct 900 gtgtcctcgg cgcaagcccg tatcgcggca cgtaaggcac gagagttggt gcggcgtaag 960 agcgccaccg acatcggtgg attgcccggc aagctggccg attgccgttc cacggatccg 1020 cgcaagtccg aactgtatgt cgtagaaggt gactcggccg gcggttctgc aaaaagcggt 1080 cgcgattcga tgttccaggc gatacttccg ctgcgcggca agatcatcaa tgtggagaaa 1140 gcgcgcatcg accgggtgct aaagaacacc gaagttcagg cgatcatcac ggcgctgggc 1200 accgggatcc acgacgagtt cgatatcggc aagctgcgct accacaagat cgtgctg 1257

[0050] <210> 15 <211> 1257 <212> DNA <213> Mycobacterium africanum <400> 15 tcggacgcgt atgcgatatc tggtggtctg cacggcgtcg gcgtgtcggt ggttaacgcg 60 ctatccaccc ggctcgaagt cgagatcaag cgcgacgggt acgagtggtc tcaggtttat 120 gagaagtcgg aacccctggg cctcaagcaa ggggcgccga ccaagaagac ggggtcaacg 180 gtgcggttct gggccgaccc cgctgttttc gaaaccacgg aatacgactt cgaaaccgtc 240 gcccgccggc tgcaagagat ggcgttcctc aacaaggggc tgaccatcaa cctgaccgac 300 gagagggtga cccaagacga ggtcgtcgac gaagtggtca gcgacgtcgc cgaggcgccg 360 aagtcggcaa gtgaacgcgc agccgaatcc actgcaccgc acaaagttaa gagccgcacc 420 tttcactatc cgggtggcct ggtggacttc gtgaaacaca tcaaccgcac caagaacgcg 480 attcatagca gcatcgtgga cttttccggc aagggcaccg ggcacgaggt ggagatcgcg 540 atgcaatgga acgccgggta ttcggagtcg gtgcacacct tcgccaacac catcaacacc 600 cacgagggcg gcacccacga agagggcttc cgcagcgcgc tgacgtcggt ggtgaacaag 660 tacgccaagg accgcaagct actgaaggac aaggacccca acctcaccgg tgacgatatc 720 cgggaaggcc tggccgctgt gatctcggtg aaggtcagcg aaccgcagtt cgagggccag 780 acc aagacca agttgggcaa caccgaggtc aaatcgtttg tgcagaaggt ctgtaacgaa 840 cagctgaccc actggtttga agccaacccc accgactcga aagtcgttgt gaacaaggct 900 gtgtcctcgg cgcaagcccg tatcgcggca cgtaaggcac gagagttggt gcggcgtaag 960 agcgccaccg acatcggtgg attgcccggc aagctggccg attgccgttc cacggatccg 1020 cgcaagtccg aactgtatgt cgtagaaggt gactcggccg gcggttctgc aaaaagcggt 1080 cgcgattcga tgttccaggc gatacttccg ctgcgcggca agatcatcaa tgtggagaaa 1140 gcgcgcatcg accgggtgct aaagaacacc gaagttcagg cgatcatcac ggcgctgggc 1200 accgggatcc acgacgagtt cgatatcggc aagctgcgct accacaagat cgtgctg 1257

<210> 16 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium avium <400> 16 gcagacgcca aagt 14

<210> 17 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium intracellulare <400> 17 cggacgccaa ggt 13

<210> 18 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium intracellulare <400> 18 ccaaggtggt ggt 13

<210> 19 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium kansasii <400> 19 gctgacgcta aaac 14

<210> 20 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium asiaticum, M.marinum and / or M. szulgai <400> 20 tcggaagcca aaac 14

<210> 21 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium asiaticum and / or M. gord onae <400> 21 ccaatccgtc gga 13

<210> 22 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium asiaticum <400> 22 gccaaaaccg ttgt 14

<210> 23 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium gastri <400> 23 gctgatgcta aaacc 15

<210> 24 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium gordonae and / or M. simia e <400> 24 tcggaagcta aaacc 15

<210> 25 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium gordonae <400> 25 gctaaaaccg ttgtg 15

<210> 26 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium malmoense <400> 26 cggatgccaa aact 14

<210> 27 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium malmoense <400> 27 gccaaaactg ttgta 15

<210> 28 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium marinum <400> 28 gccaatcctt cgga 14

<210> 29 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium marinum <400> 29 gccaaaacca ttgtg 15

<210> 30 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium scroflaceum <400> 30 tccgaggcga aaac 14

<210> 31 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium scroflaceum <400> 31 ccaacccatc gga 13

<210> 32 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium scroflaceum <400> 32 cgaaaacggt ggtg 14

<210> 33 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium simiae <400> 33 gctaaaaccg ttgta 15

<210> 34 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium szulgai <400> 34 ccaacccgtc gga 13

<210> 35 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium szulgai <400> 35 ccaaaaccgt cgtg 14

<210> 36 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium tuberculosis <400> 36 accgacgcga aagt 14

<210> 37 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium tuberculosis <400> 37 ccaaccccac cga 13

<210> 38 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium tuberculosis and / or M.g astri <400> 38 aggtgtgcaa tgaac 15

<210> 39 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium bovis <400> 39 tctgtaatga acagc 15

<210> 40 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium bovis <400> 40 aggtctgtaa cgaac 15

<210> 41 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium microti, M. africanum and / or M. bovis <400> 41 accgactcga aagt 14

<210> 42 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium microti, M. africanum and / or M. tuberculosis <400> 42 tctgtaacga acagc 15

<210> 43 <211> 14 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid for detecting Mycobacterium microti, M. africanum, M. Tuberculosis and / or M. bovis <400> 43 gcgaaagtcg ttgt 14

<210> 44 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid used for amplifying a region in gyrB gene of Mycobacterium <400> 44 tgtaaaacga cggccagtca ygcnggnggn aarttyga 38

<210> 45 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid used for amplifying a region in gyrB gene of Mycobacterium <400> 45 ctgcgttcgt atatgagcnc crtcnacrtc ngcrtc 36

<210> 46 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid used for amplifying a region in gyrB gene of Mycobacterium <400> 46 cayctcnggn ggnaarttyg a 21

<210> 47 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Nucleic Acid used for amplifying a region in gyrB gene of Mycobacterium <400> 47 tcnacrtcng crtcngtcat 20

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) (C12Q 1/68 C12R 1:32) C12R 1:32) C12N 15/00 ZNAA (72) Inventor Masao Fukushima 51 Komiyacho, Hachioji-shi, Tokyo SRL Hachioji Laboratory Co., Ltd. (72) Inventor Kenichi Kakinuma 51 Komiyacho, Hachioji-shi, Tokyo Inside SRL Hachioji Laboratory Co., Ltd. (72) Ryuuji Kawaguchi 51 Komiyacho, Hachioji-shi, Tokyo Stock (72) Inventor Hiroaki Kasai 75-1 Hirata No.3, Hirata, Kamaishi City, Iwate Prefecture F-term (reference) 4K024 AA13 CA09 GA19 HA14 4B063 QA01 QA18 QQ06 QQ42 QR32 QR39 QR56 QR62 QR84 QS03 QS25 QS34

Claims (71)

[Claims] 1. A region consisting of 8 or more contiguous nucleotides in the 867 nt to 900 nt region of the nucleotide sequence represented by SEQ ID NO: 1 in the sequence listing, wherein at least one of g of 877 nt, a of 879 nt, and c of 885 nt A nucleic acid for detecting Mycobacterium avium, comprising a nucleic acid fragment having a base sequence of a region containing: 2. The nucleic acid for detecting Mycobacterium avium according to claim 1, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 16 in the sequence listing. 3. A region of 8 or more contiguous bases in the region of 868 nt to 900 nt of the base sequence represented by SEQ ID NO: 2 in the sequence listing, wherein at least one of g of 879 nt, c of 885 nt, and g of 888 nt A nucleic acid for detecting Mycobacterium intracellulare comprising a nucleic acid fragment comprising a region containing: 4. The nucleic acid for detecting Mycobacterium intracellulare according to claim 3, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 17 in the sequence listing. 5. A continuous region of 8 bases or more in the 874 nt to 906 nt region of the base sequence represented by SEQ ID NO: 2 in the sequence listing, wherein at least one of g of 888 nt, g of 891 nt, and g of 894 nt A nucleic acid for detecting Mycobacterium intracellulare comprising a nucleic acid fragment comprising a region containing: 6. The nucleic acid for detecting Mycobacterium intracellulare according to claim 5, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 18 in the sequence listing. 7. A region of 8 or more consecutive nucleotides in the 861 nt to 904 nt region of the nucleotide sequence represented by SEQ ID NO: 3 in the sequence listing, wherein 871 nt, 873 nt, 879 nt and 883 nt a A nucleic acid for detecting Mycobacterium kansasii comprising a nucleic acid fragment comprising a region containing at least one of the following: 8. The nucleic acid for detecting Mycobacterium kansasii according to claim 7, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 19 in the sequence listing. 9. A nucleic acid for detecting Mycobacterium asiaticum, Mycobacterium marinum and / or Mycobacterium sulgai, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 20 in the sequence listing. 10. A region in SEQ ID NO: 4, 8 or 11 of the sequence listing, wherein the region extends from 10 bases upstream to 10 bases downstream of the region having the base sequence represented by SEQ ID NO: 20 in the sequence listing. It is a continuous region of 8 or more bases, wherein 1 nt t, 3 nt g, 6n
Mycobacterium asiaticum, Mycobacterium marinum and / or Mycobacterium comprising a nucleic acid fragment having a nucleotide sequence of a region containing at least one of t a, 9 nt c, 13 nt a and 14 nt c -Nucleic acid for detecting squid 11. A nucleic acid for detecting Mycobacterium asiaticum and / or Mycobacterium gordonae comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 21 in the sequence listing. 12. A region in the sequence listing of SEQ ID NO: 4 or 6, which is continuous in a region ranging from 10 bases upstream to 10 bases downstream from the region having the base sequence represented by SEQ ID NO: 21 in the sequence listing. It consists of a nucleic acid fragment having a base sequence of a region having at least 8 bases and including at least one of 5 nt t, 8 nt g, 9 nt t and 11 nt g in the base sequence represented by SEQ ID NO: 21. A nucleic acid for detecting Mycobacterium asiaticum and / or Mycobacterium gondae. 13. A continuous region of 8 or more bases in the region from 873 nt to 906 nt of the base sequence represented by SEQ ID NO: 4 in the sequence listing, wherein at least one of c of 885 nt, a of 889 nt, and c of 890 nt A nucleic acid for detecting Mycobacterium asiaticum comprising a nucleic acid fragment comprising a region containing: 14. The nucleic acid for detecting Mycobacterium asiaticum according to claim 13, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 22 in the sequence listing. 15. A region of 8 or more consecutive bases in the 861 nt to 895 nt region of the base sequence represented by SEQ ID NO: 5 in the sequence listing, wherein 871 nt g, 873 nt t, 876 nt t, 876 nt t, and 879 nt t ,
A nucleic acid for detecting Mycobacterium gastori comprising a nucleic acid fragment comprising a region containing at least one of 883 nt a and 884 nt c. 16. The nucleic acid for detecting Mycobacterium gastri according to claim 15, comprising a nucleic acid fragment having the base sequence represented by SEQ ID NO: 23 in the sequence listing. 17. A nucleic acid for detecting Mycobacterium gordonae and / or Mycobacterium simiae comprising a nucleic acid fragment having the nucleotide sequence of SEQ ID NO: 24 in the sequence listing. 18. A region in SEQ ID NO: 6 or 10 in the sequence listing, which is continuous in a region ranging from 10 bases upstream to 10 bases downstream from the region having the base sequence represented by SEQ ID NO: 24 in the sequence listing. It is a region of 8 bases or more, in the base sequence represented by SEQ ID NO: 24, 1 nt t, 3 nt g, 6 nt
a, a nucleic acid for detecting Mycobacterium gordonae and / or Mycobacterium simiae comprising a nucleic acid fragment having a nucleotide sequence of a region containing at least one of a, 13 nt a and 14 nt c. 19. A continuous region of not less than 8 bases in the 873 nt to 907 nt region of the nucleotide sequence represented by SEQ ID NO: 6 in the sequence listing, wherein at least one of t of 885 nt, a of 889 nt, and c of 890 nt A nucleic acid for detecting Mycobacterium gondae, comprising a nucleic acid fragment comprising a region comprising: 20. The nucleic acid for detecting Mycobacterium gordonae according to claim 19, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 25 in the sequence listing. 21. A continuous region of 8 bases or more in the region from 868 nt to 901 nt of the base sequence represented by SEQ ID NO: 7 in the sequence listing, wherein 879 nt is g, 882 nt is t, 885 nt is c, and 889 nt is a , 8
A nucleic acid for detecting Mycobacterium marmoense comprising a nucleic acid fragment comprising a region containing at least one of 90 nt c and 891 nt t. 22. The nucleic acid for detecting Mycobacterium marmoense according to claim 21, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 26 in the sequence listing. 23. A continuous region of 8 or more bases in the 872 nt to 907 nt region of the base sequence represented by SEQ ID NO: 7 in the sequence listing, wherein 885 nt c, 889 nt a, 890 nt c and 891 nt t
A nucleic acid for detecting Mycobacterium marmoense comprising a nucleic acid fragment comprising a region containing at least one of the following. 24. The nucleic acid for detecting Mycobacterium marmoense according to claim 23, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 27 in the sequence listing. 25. A continuous region of not less than 8 bases in the region from 858 nt to 881 nt of the nucleotide sequence represented by SEQ ID NO: 8 in the sequence listing, wherein t is 873 nt, t is 876 nt, t is 877 nt, and g is 879 nt.
A nucleic acid for detecting Mycobacterium marinum, comprising a nucleic acid fragment comprising a region containing at least one of the following. 26. The nucleic acid for detecting Mycobacterium marinum according to claim 25, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 28 in the sequence listing. 27. A continuous region of not less than 8 bases in the 873 nt to 907 nt region of the base sequence represented by SEQ ID NO: 8 in the sequence listing, wherein 885 nt c, 889 nt a, 890 nt c and 892 nt a
A nucleic acid for detecting Mycobacterium marinum, comprising a nucleic acid fragment comprising a region containing at least one of the following. 28. The nucleic acid for detecting Mycobacterium marinum according to claim 27, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 29 in the sequence listing. 29. A continuous region of 8 or more bases in the region of 867 nt to 900 nt of the base sequence represented by SEQ ID NO: 9 in the sequence listing, wherein t is 877 nt, g is 882 nt, g is 889 nt, and c is 890 nt.
A nucleic acid for detecting Mycobacterium scroflaseum comprising a nucleic acid fragment comprising a region containing at least one of the following. 30. The nucleic acid for detecting Mycobacterium scroflaseum according to claim 29, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 30 in the sequence listing. 31. A region consisting of at least 8 consecutive nucleotides in the 858 nt to 891 nt region of the nucleotide sequence represented by SEQ ID NO: 9 in the sequence listing, and containing at least one of a 876 nt a and a 877 nt t For detecting Mycobacterium scroflaseum comprising a nucleic acid fragment comprising: 32. The nucleic acid for detecting Mycobacterium scroflaseum according to claim 31, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 31 in the sequence listing. 33. A region of at least 8 contiguous bases in the 874 nt to 907 nt region of the base sequence represented by SEQ ID NO: 9 in the sequence listing, wherein the region includes at least any one of 889 nt a to 891 nt g. For detecting Mycobacterium scroflaseum comprising a nucleic acid fragment comprising: 34. The nucleic acid for detecting Mycobacterium scroflaseum according to claim 33, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 32 in the sequence listing. 35. A continuous region of not less than 8 bases in the 873 nt to 907 nt region of the base sequence represented by SEQ ID NO: 10 in the sequence listing, wherein 885 nt t, 889 nt a, 890 nt c, and 897 nt a
A nucleic acid for detecting Mycobacterium simiae comprising a nucleic acid fragment comprising a region containing at least one of the following. 36. The nucleic acid for detecting Mycobacterium simiae according to claim 35, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 33 in the sequence listing. 37. A region consisting of 8 or more contiguous nucleotides in the 859 nt to 891 nt region of the nucleotide sequence represented by SEQ ID NO: 11 in the sequence listing, wherein at least one of g of 876 nt, t of 877 nt, and g of 879 nt A nucleic acid for detecting Mycobacterium surugai, comprising a nucleic acid fragment comprising a region containing 38. The nucleic acid for detecting Mycobacterium surugai according to claim 37, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 34 in the sequence listing. 39. A continuous region of 8 bases or more in the 874 nt to 907 nt region of the base sequence represented by SEQ ID NO: 11 in the sequence listing, wherein 885 nt c, 889 nt a, 890 nt c and 894 nt
A nucleic acid for detecting Mycobacterium surugai, comprising a nucleic acid fragment comprising a region containing at least any one of the above c. 40. The nucleic acid for detecting Mycobacterium surugai according to claim 39, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 35 in the sequence listing. 41. A nucleic acid for detecting Mycobacterium tuberculosis comprising a nucleic acid fragment comprising a continuous region of 8 or more bases in a region of 861 nt to 894 nt of the base sequence represented by SEQ ID NO: 12 in the sequence listing. 42. The nucleic acid for detecting Mycobacterium tuberculosis according to claim 41, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 36 in the sequence listing. 43. A nucleic acid for detecting Mycobacterium tuberculosis comprising a nucleic acid fragment comprising a continuous region of 8 or more bases in the region of 853 nt to 885 nt of the base sequence represented by SEQ ID NO: 12 in the sequence listing. 44. The nucleic acid for detecting Mycobacterium tuberculosis according to claim 43, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 37 in the sequence listing. 45. A continuous eight or more bases region in the region of 817 nt to 851 nt of the base sequence represented by SEQ ID NO: 5 in the sequence listing, wherein at least one of g of 831 nt, c of 834 nt, and t of 837 nt A nucleic acid for detecting Mycobacterium gastori comprising a nucleic acid fragment comprising a region comprising: 46. The nucleic acid for detecting Mycobacterium gastri according to claim 45, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 38 in the sequence listing. 47. A mycobacterium comprising a nucleic acid fragment consisting of a continuous region of at least 8 bases in the 820 nt to 854 nt region of the base sequence represented by SEQ ID NO: 13 in the sequence listing and comprising at least 837 nt t. Umbobis detection nucleic acid. 48. The nucleic acid for detecting Mycobacterium bovis according to claim 47, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 39 in the sequence listing. 49. A nucleic acid for detecting Mycobacterium bovis, Mycobacterium microti and / or Mycobacterium africanum, comprising a nucleic acid fragment having the nucleotide sequence represented by SEQ ID NO: 41 in the sequence listing. 50. SEQ ID NO: 13, 14 or 15 in the sequence listing
A region having a base sequence represented by SEQ ID NO: 41 in the sequence listing, a continuous region of 8 or more bases in a region extending from 10 bases upstream to 10 bases downstream from the region having the base sequence represented by SEQ ID NO: 41, A nucleic acid for detecting Mycobacterium bovis, Mycobacterium microti and / or Mycobacterium africanum, comprising a nucleic acid fragment having a nucleotide sequence of a region containing at least 7 nt of t in the nucleotide sequence shown. 51. For detecting Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium microti and / or Mycobacterium africa nam consisting of a nucleic acid fragment having the nucleotide sequence shown by SEQ ID NO: 42 in the sequence listing. Nucleic acids. 52. SEQ ID NO: 12, 14, or 15 in the sequence listing
From a nucleic acid fragment having a base sequence of a continuous 8 or more bases region in a region ranging from 10 bases upstream to 10 bases downstream of the region having the base sequence represented by SEQ ID NO: 42 in the sequence listing. A nucleic acid for detecting Mycobacterium tuberculosis, Mycobacterium microti and / or Mycobacterium africanum. 53. For detecting Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium microti and / or Mycobacterium africanum consisting of a nucleic acid fragment having the nucleotide sequence represented by SEQ ID NO: 43 in the sequence listing. Nucleic acids. 54. A region in SEQ ID NO: 12, 13, 14 or 15 in the sequence listing, wherein the region extends from 10 bases upstream to 10 bases downstream of the region having the base sequence represented by SEQ ID NO: 43 in the sequence listing. A nucleic acid for detecting Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium microti and / or Mycobacterium africanum, comprising a nucleic acid fragment having a base sequence of a continuous 8 or more base region therein. 55. A nucleic acid for detecting Mycobacterium bovis comprising a nucleic acid fragment comprising a continuous region of 8 bases or more in the region of 817 nt to 851 nt of the base sequence represented by SEQ ID NO: 13 in the sequence listing. 56. The nucleic acid for detecting Mycobacterium bovis according to claim 55, comprising a nucleic acid fragment having a base sequence represented by SEQ ID NO: 40 in the sequence listing. 57. The nucleic acid according to any one of claims 1 to 56, wherein the nucleic acid has 8 to 50 bases. 58. The nucleic acid according to any one of claims 1 to 57, which is a probe. 59. A method for detecting a bacterium belonging to the genus Mycobacterium, comprising using a plurality of the nucleic acids for detection according to claim 1 to 58. 60. Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium kansasii, Mycobacterium asiaticum, Mycobacterium gastri, Mycobacterium gordonae, Mycobacterium marmoense ,
Mycobacterium marinum, Mycobacterium
Of the fifteen Mycobacterium bacteria, Scrofraceum, Mycobacterium simiae, Mycobacterium surugai, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium microti and Mycobacterium africanum, 7
60. The method of claim 59, wherein more than one species is detectably detectable. 61. The method according to claim 60, wherein 10 or more species can be detected differentially. 62. The method according to claim 60, wherein the method is carried out using seven or more types of nucleic acids having the nucleotide sequences of SEQ ID NOs: 16 to 43 in the sequence listing. 63. The method according to claim 61, wherein the method is carried out using 10 or more types of nucleic acids having the nucleotide sequences of SEQ ID NOs: 16 to 43 in the sequence listing. 64. The method according to claim 63, wherein the method is carried out using 15 or more types of nucleic acids having the nucleotide sequences of SEQ ID NOs: 16 to 43 in the sequence listing. 65. A kit for detecting a bacterium belonging to the genus Mycobacterium, comprising a plurality of the nucleic acids for detection according to claim 1 to 58. 66. Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium kansasii, Mycobacterium asiaticum, Mycobacterium gastri, Mycobacterium gordonae, Mycobacterium marmoense ,
Mycobacterium marinum, Mycobacterium
Of the fifteen Mycobacterium bacteria, 15 species of Scrofraceum, Mycobacterium simiae, Mycobacterium surugai, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium microti and Mycobacterium africanum
67. The kit of claim 66, wherein more than one species can be differentially detected. 67. The kit according to claim 66, wherein 10 or more species can be detected differentially. 68. The kit according to claim 66, comprising seven or more nucleic acids having the nucleotide sequences of SEQ ID NOs: 16 to 43 in the sequence listing. 69. The method according to claim 67, comprising 10 or more nucleic acids having the nucleotide sequences of SEQ ID NOs: 16 to 43 in the sequence listing.
The kit as described. 70. A nucleic acid comprising 15 or more nucleic acids having the nucleotide sequences of SEQ ID NOs: 16 to 43 in the sequence listing.
The kit as described. The kit according to any one of claims 65 to 70, wherein a plurality of types of nucleic acids are carried on one carrier.