JPH0361487A - Detection of swine epidemic pneumonia mycoplasma - Google Patents
Detection of swine epidemic pneumonia mycoplasmaInfo
- Publication number
- JPH0361487A JPH0361487A JP1198475A JP19847589A JPH0361487A JP H0361487 A JPH0361487 A JP H0361487A JP 1198475 A JP1198475 A JP 1198475A JP 19847589 A JP19847589 A JP 19847589A JP H0361487 A JPH0361487 A JP H0361487A
- Authority
- JP
- Japan
- Prior art keywords
- dna
- base sequence
- probe
- formula
- nucleotide base
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
- 241000282898 Sus scrofa Species 0.000 title claims abstract description 9
- 206010035664 Pneumonia Diseases 0.000 title claims abstract description 8
- 241000204031 Mycoplasma Species 0.000 title claims abstract 6
- 238000001514 detection method Methods 0.000 title description 7
- 239000000523 sample Substances 0.000 claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 21
- 239000002773 nucleotide Substances 0.000 claims abstract description 8
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 8
- 238000009396 hybridization Methods 0.000 claims abstract description 7
- 108020004414 DNA Proteins 0.000 claims description 10
- 241000204045 Mycoplasma hyopneumoniae Species 0.000 claims description 4
- 108020004511 Recombinant DNA Proteins 0.000 claims description 2
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- 241000894006 Bacteria Species 0.000 description 3
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 108020003215 DNA Probes Proteins 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000002405 diagnostic procedure Methods 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 1
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 1
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- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
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Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は豚流行性肺炎原因菌マイコプラズマ・ハイオニ
ュウモニエの検出方法に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for detecting Mycoplasma hyopneumoniae, a bacterium that causes swine epidemic pneumonia.
豚流行性肺炎（豚マイコプラズマ肺炎Ｍｙｃｏｐｌａｓ
ｍａＰｎｅｕｍｏｎｉａｅ ｏｆ ５ｗ１ｎｅ＋以下ｒ
ＭＰｓＪという）はマイコプラズマ・ハイオニュウモニ
エ（Ｍｙｃｏｐｌａｓｍａｈｙｏｐｎｅｕｍｏｎｉａｅ
） （以下、ｒＭ、ｈｐＪという）という微生物により
惹起される。本店は慢性経過を取るのが特徴で、死亡率
は低い。しかしながら罹患率は極めて高く、飼料効率の
低下等、養豚産業に与えている経済的損失は美大なもの
となっている。Porcine epidemic pneumonia (Mycoplas pneumonia)
maPneumoniae of 5w1ne+r below
MPsJ) is Mycoplasma hyopneumoniae.
) (hereinafter referred to as rM, hpJ). The main disease is characterized by a chronic course, and the mortality rate is low. However, the morbidity rate is extremely high, and the economic losses caused to the pig farming industry, such as decreased feed efficiency, are enormous.
農林水産省の調査によると現在日本の層場に出荷される
豚の半数以上が本店による肺病変を有しており、はぼ全
国的に蔓延していることが明らかにされている。According to a survey by the Ministry of Agriculture, Forestry and Fisheries, more than half of the pigs shipped to Japan's stockyards now have lung lesions caused by this disease, and it has become clear that the disease is widespread nationwide.
ＭＰＳの診断法に関しては補体結合反応、ＢＬＩＳＡ法
等による血清診断法、あるいは肺病変の肉眼所見による
方法が報告されている。また、病巣からＭ、ｈｐを分離
する方法もあるが、Ｍ、ｈｐは栄養要求性が厳しく人工
培地での発育には長期間が必要である。このため、従来
Ｍｐ ｓ ｇ染豚からＭ、ｈｐを分離し同定するまでに
少なくとも２週間から１ケ月程度必要であった。Regarding diagnostic methods for MPS, a serum diagnostic method using complement fixation reaction, BLISA method, etc., or a method based on macroscopic findings of lung lesions has been reported. There is also a method of separating M and hp from lesions, but M and hp have strict nutritional requirements and require a long period of time to grow in an artificial medium. For this reason, it conventionally required at least two weeks to one month to isolate and identify M and hp from Mpsg-stained pigs.
本発明の目的は、病巣からＭ、ｈｐを短期間で検出する
方法を提供することである。An object of the present invention is to provide a method for detecting M, hp from lesions in a short period of time.
本発明の目的は以下の式（Ｉ）、（ｎ）、（Ｉ）、（Ｉ
Ｖ）または（Ｖ）で示される塩基配列のヌクレオチド、
これらをその一部として含む組換えＤＮＡ。The object of the present invention is to have the following formulas (I), (n), (I), (I
V) or nucleotides of the base sequence shown by (V),
Recombinant DNA containing these as part of it.
あるいはこれらをその一部として含む合ａＤＮＡをＤＮ
Ａプローブとして用いたＤＮＡハイブリダイゼーション
法により達成される。Or DNA containing these as part of DNA
This is achieved by a DNA hybridization method using an A probe.
ししＩｊＬＴＡＡＬＵ’１ Ｕｉ’ｌｔＡ’ｌ’ＡＡ’１ｒ ＵＴＡＴＴＴＴＧ（：［’ 以下、本発明の詳細な説明する。Shishi IjLTAALU’1 Ui’ltA’l’AA’1r UTATTTTTG(:[’ The present invention will be explained in detail below.
リボゾームＲＮＡ （以下ｒ ｒＲＮＡ」という）は１
菌体当り１万コピー以上存在し、検出感度の面では非常
に有効であると考えられる。しかし、ｒ ＲＮ Ａは生
物の進化の過程において比較的よく保存されたものの一
つであることから抗原遺伝子と比較して特異性が低い。Ribosomal RNA (hereinafter referred to as r rRNA) is 1
There are over 10,000 copies per bacterial cell, and it is considered to be very effective in terms of detection sensitivity. However, since rRNA is one of the genes that has been relatively well conserved during the evolution of living organisms, its specificity is lower than that of antigen genes.
たとえば大腸菌、と枯草菌では７０＝８０％相同性があ
ると言われている。For example, Escherichia coli and Bacillus subtilis are said to have 70=80% homology.
ところでＭ、ｈｐのｒＲＮＡについては、西独のグルー
プが１６５ｒＲＮＡの塩基配列を決定している〔Ｃ，Ｔ
ａ５ｃｈｋｅ ｅｔ ａｌ、 Ｎｕｃｌ、 Ａｃ１ｄｓ
Ｒｅ５１１５、３９１８（１９８７） ”Ｎｕｃｌｅ
ｏｔｉｄｅ ５ｅｑｕｅｎｃｅ ｏｆ ｔｈｅ１６Ｓ
ｒＲＮＡ ｏｆ Ｍｙｃｏｐｌａｓｍａ ｈｙｏｐｎｅ
ｕｍｏｎｉａｅ” ：ｌ 。本発明者は、このデータを
用い、現在までにすでに決定されている他種生物の１６
ＳｒＲＮＡの塩基配列と比較してＭ、ｈｐ １６Ｓ ｒ
ＲＮＡに特有な領域を検索した。By the way, regarding the rRNA of M and hp, a group in West Germany has determined the base sequence of 165 rRNA [C, T
a5chke et al, Nucl, Ac1ds
Re5115, 3918 (1987) “Nucle
otide 5equence of the16S
rRNA of Mycoplasma hyopne
Umoniae": l. Using this data, the present inventors have identified the 16 species of other organisms that have been determined to date.
Compared with the base sequence of SrRNA, M, hp 16S r
We searched for regions unique to RNA.
その結果、Ｍ、ｈｐに特異的と考えられる数個の領域を
発見し、その領域からＤＮＡプローブとして有用と考え
られるオリゴヌクレオチドを選定し、これを化学合成し
、ＤＮＡプローブとしてＩＶｌｈｐの検出を行ったとこ
ろ、極めて高感度でＭ、ｈｐを検出しうろことを見出し
、本発明を完成するに至った。As a result, we discovered several regions thought to be specific to M, hp, selected oligonucleotides considered to be useful as DNA probes from these regions, chemically synthesized them, and detected IVlhp as DNA probes. As a result, they discovered that scales can detect M and hp with extremely high sensitivity, and have completed the present invention.
（、へ） Ｍ、 ｈｐ １６ Ｓ ｒＲＮＡにおけ
る特異領域の検索
ＪＯＩＳ−ＦのＧｅｎ １３ａｎｋ及びＥＭＢＬＤＮＡ
データベースを用い現在までに報告されている各種生物
の１６３ｒＲＮＡの塩基配列を検索した。(, to) M, Search for unique regions in hp 16 S rRNA JOIS-F Gen 13ank and EMBL DNA
Using a database, we searched for 163 rRNA base sequences of various organisms that have been reported to date.
得られた塩基配列すべてと、第１図に示したＩｖｆ、ｈ
ｐ １６ Ｓ ｒＲＮＡの塩基配列とを、遺伝子解析
用パソコン・ソフトウェアＧＥＮＩＡＳ （三井情報開
発）を用いて比較した。All the obtained base sequences and Ivf, h shown in Figure 1
The base sequence of p 16 S rRNA was compared using computer software for gene analysis GENIAS (Mitsui Knowledge Development).
その結果、第１図において、塩基Ｎｏ、 １８０〜２４
０．５９０〜６８０．９９０−１０４０の３ケ所がＭ、
ｈｐ特異領域として考えられることがわかった。As a result, in FIG. 1, base No. 180-24
3 places from 0.590 to 680.990-1040 are M,
It was found that this region can be considered as a hp-specific region.
（Ｂ） オリゴヌクレオチドの合成
法に、ＤＮＡプローブとして有用と考えられる、化学合
成すべきオリゴヌクレオチドを、以下の点に注意して決
定した。(B) Oligonucleotides to be chemically synthesized that are considered useful as DNA probes in the oligonucleotide synthesis method were determined with the following points in mind.
〔１）分子内で高次構造をとらないこと。[1) Do not form a higher-order structure within the molecule.
（２） １６ＳｒＲＮＡの他の領域と相同性をもたな
いこと。(2) It has no homology with other regions of 16S rRNA.
（３）分子内にくり返し構造をもたないこと。(3) No repeating structure within the molecule.
（４）塩基の組成比が極端にかたよらないこと。(4) The composition ratio of the base should not vary significantly.
このような条件を満たし、ＤＮＡプローブとして適当と
考えられる３０塩基長のオリゴヌクレオチド２種類（プ
ローブ１及び２）を台底した。プローブ１及び２の塩基
配列は前記式（ＩＶ）及び（Ｖ）により表わされる。な
お、このオリゴヌクレオチドは１６Ｓ ｒＲＮＡの逆相
補鎖である。すなわち、プローブ１はＭ、ｈｐ １６５
ｒＲＮＡの塩基Ｎｏ、 ２０３〜２３２、またプロー
ブ２は塩基Ｎｏ、 ６１０〜６３９の逆相補鎖にあたる
。Two types of oligonucleotides (probes 1 and 2) with a length of 30 bases that meet these conditions and are considered suitable as DNA probes were developed. The base sequences of probes 1 and 2 are represented by formulas (IV) and (V) above. Note that this oligonucleotide is a reverse complementary strand of 16S rRNA. That is, probe 1 is M, hp 165
rRNA base numbers 203-232, and probe 2 corresponds to the reverse complementary strand of base numbers 610-639.
（Ｃ）”Ｐ標識合成オリゴヌクレオチドを用いたＤＮＡ
プローブ法によるＭ、ｈｐの検出合成オリゴヌクレオチ
ドの５′末端を３２ｐで標識し、Ｍ、ｈｐのＤＮＡプロ
ーブ法による検出を試みた。(C) “DNA using P-labeled synthetic oligonucleotide
Detection of M and hp by probe method The 5' end of a synthetic oligonucleotide was labeled with 32p, and detection of M and hp by DNA probe method was attempted.
１） ” Ｐ標識合成オリゴヌクレオチドの調製Ｔ４ポ
リヌクレオチドキナーゼ及びｒｒ−”Ｐｌ ＡＴＰを用
い、台底オリゴヌクレオチドの５′末端をリン酸化する
ことにより３２ｐｇ識を行った。得られたオリゴヌクレ
オチドの比放射活性は約４ Ｘ １０’ ｃ、ｐ、ｍ、
／ｐｍｏｌｅであった。1) Preparation of ``P-labeled synthetic oligonucleotide 32pg labeling was performed by phosphorylating the 5' end of the platform oligonucleotide using T4 polynucleotide kinase and rr-''Pl ATP. The specific radioactivity of the obtained oligonucleotide was approximately 4 X 10' c,p,m,
/pmole.
２）ＤＮＡプローブ法
上記プローブはｒＲＮＡに結合するものであす、シかも
オリゴヌクレオチドプローブであることから、−船釣な
ハイブリダイゼーションの条件ではうまく反応がおこら
ない。2) DNA probe method Since the above probe binds to rRNA and is also an oligonucleotide probe, the reaction does not occur under harsh hybridization conditions.
そこで培養液の処理法、ハイブリダイゼーションの方法
、メンブレンの選択等、種々の条件検討を行った。Therefore, we investigated various conditions such as culture solution treatment methods, hybridization methods, and membrane selection.
その結果、次に示すような方法が適当であることが明ら
かとなった。As a result, it became clear that the following method was appropriate.
被検液１００μｌに２０％ジチオスレイトール又は２０
％Ｎ−アセチル−Ｌ−システィン１００μｌを加え、３
７℃、３０分処理し、溶菌させる。Add 20% dithiothreitol or 20% dithiothreitol to 100 μl of test solution.
Add 100 μl of %N-acetyl-L-cysteine and
Treat at 7°C for 30 minutes to lyse.
この処理により溶菌のほか、豚の鼻汁等の粘度の高い試
料を低粘度化できる。この試料をメンブレン（ゲルマン
社製バイオトレースＲＰ等）上にスポットする。８０℃
で２時間乾燥後、６０℃のプレハイブリダイゼーション
溶液（： ５ Ｘ５ＳＣ（０，１５ＭＮａＣ１，０，０
１５Ｍクエン酸ナトリウム〉、５ ｘＤｅｎｈａｌｄｔ
’ Ｓ溶液（０，０２％ＢＳＡ、０．０２％ポリビニル
ピロリドン、０．０２％フィコール）、０．２％５ＤＳ
（ドデシル硫酸ナトリウム）、２００μｇ／ｍｌ酵母ｔ
ＲＮ Ａ ：］に１時間浸漬する。続いて６０℃のハ
イブリダイゼーション溶液〔ブレハイブリダイゼーショ
ン溶液に２ ×１０ｇｃ、ｐ、ｍ、 ７１０ｍＩ！の３
２Ｐ標ｍオリゴヌクレオチドを加えたもの〕に２時間浸
漬する。In addition to lysing bacteria, this treatment can reduce the viscosity of highly viscous samples such as pig nasal secretions. This sample is spotted onto a membrane (Biotrace RP manufactured by Gelman, etc.). 80℃
After drying for 2 hours at 60 °C, prehybridization solution (: 5
15M Sodium Citrate>, 5 x Denholdt
'S solution (0.02% BSA, 0.02% polyvinylpyrrolidone, 0.02% Ficoll), 0.2% 5DS
(sodium dodecyl sulfate), 200 μg/ml yeast t
Soak in RNA:] for 1 hour. Next, add 60°C hybridization solution [2 x 10gc, p, m, 710mI! No. 3
2P-marked oligonucleotide] for 2 hours.
６０℃の５ｘＳＳＣ−０，２％ＳＤＳ溶液で２分間づつ
、２回洗浄し、風乾後、Ｘ線フィルム及び増感スクリー
ンを用いて、−７０℃で１６時間オートラジオグラフィ
ーを行う。After washing twice with 5xSSC-0.2% SDS solution at 60°C for 2 minutes each time and air drying, autoradiography is performed at -70°C for 16 hours using an X-ray film and an intensifying screen.
Ｍ、ｈｐとＭ、ｈｒ （Ｍｙｃｏｐｌａｓ＋ｎａ ｈｙ
ｏｒｈｉｎｉｓ）を２日間培養したものを原液とし、こ
の原液を試料１、その５．１０．５０．１００及び１０
００倍希釈液をそれぞれ試料２．３．４．５及び６とし
、その各１００μｍについて、上記方法にしたがって、
プローブ１及び２を用いてＭ、ｈｐの検出を行った。結
果を第２図に示す。（ハ）はプローブ１とプローブ２の
量をそれぞれＩ Ｘ １０６ｃ、ｐ、ｍ、／ １０−使
用したときの結果を示す。（イ）、（ロ）、（ハ）いず
れのばあいにも、１０００倍希釈液においてＭ、ｈｐを
検出することができた。Ｍ、ｈｐの培養原液の菌数は１
０６〜１０’ ｃｆｕ ／ｍ１．であリ、したがって１
０２〜１０’個の菌数で十分検出できることがわかる。M, hp and M, hr (Mycoplas+na hy
Orhinis) was cultured for 2 days and used as a stock solution, and this stock solution was used as sample 1, sample 5.10.50.
Samples 2, 3, 4, 5 and 6 were prepared using the 00 times diluted solution, and each 100 μm of the solution was prepared according to the above method.
M and hp were detected using probes 1 and 2. The results are shown in Figure 2. (c) shows the results when probe 1 and probe 2 were used in amounts of I x 106c, p, m, /10-, respectively. In all cases (a), (b), and (c), M and hp could be detected in the 1000-fold diluted solution. The number of bacteria in the stock culture solution of M and hp is 1.
06-10'cfu/m1. Therefore, 1
It can be seen that a bacterial count of 02 to 10' can be sufficiently detected.
またＭ、ｈｐに最も近縁のＭ、 ｈｒについては培養
原液でも全く反応しなかった。Furthermore, M and hr, which are most closely related to M and hp, did not react at all even with the stock culture solution.
また２種類のオリゴヌクレオチドは共にＭ、ｈｐに対す
る特異性があり、両者の混合により、さらに検出感度を
上げることができる。Furthermore, both of the two types of oligonucleotides have specificity for M and hp, and by mixing the two, detection sensitivity can be further increased.
本発明のＤＮＡをプローブとして用いるＭ、ｈｐの検出
法は、通常の分離検出法と比較して短時間（約２日）で
行うことができる。さらに、サンプルをジチオスレイト
ール又はＮ−アセチル−Ｌ−ンステイン処理するばあい
には、粘度の高い鼻汁等のサンプルについても適用可能
である。The method for detecting M and hp using the DNA of the present invention as a probe can be performed in a shorter time (about 2 days) than conventional separation detection methods. Furthermore, when the sample is treated with dithiothreitol or N-acetyl-L-steine, it can also be applied to samples such as highly viscous nasal secretions.
Ｍ、ｈｐ感染豚の鼻汁には１０’ 〜１０’ ｃｆｕ／
−のＭ、ｈｐが存在すると考えられているので鼻汁試料
を用いてＭ、ｈｐ感染豚の直接診断も可能である。The nasal secretions of M, hp-infected pigs contain 10' to 10' cfu/
- Since it is thought that M.hp.- is present, it is also possible to directly diagnose M.hp. infected pigs using nasal discharge samples.
第１図は、Ｍ、ｈｐの１６ＳｒＲＮＡの全塩基配列を示
す。
第２図は、
本発明のプローブ１及び２を用いて
行ったＭ。
ｈｐ
検出試験の結果を示す。FIG. 1 shows the entire base sequence of 16S rRNA of M and hp. FIG. 2 shows M measurements performed using probes 1 and 2 of the present invention. The results of the hp detection test are shown.
Claims (1)
るヌクレオチド塩基配列を有するＤＮＡ。 （ I ） ＣＣＣＣＡＴＴＴＣ＾１Ｔ＾０ＴＧＧＴＧＡＡＧＣ＾２
Ｔ＾０ＴＧＡＡＧＧＣＴＣ＾３Ｃ＾０ＴＴＴＧＡＡＴＡ
Ａ＾４Ａ＾０ＡＡＴＴＣＡＴＧＣ＾５Ａ＾０ＡＡＴＴＴ
ＴＴＡＴ＾６Ｃ＾０Ａ（II） ＣＣＧＣＴＡＡＣＣ＾１Ｔ＾０ＣＴＴＣＡＴＡＡＴ＾２
Ｔ＾０ＣＴＡＴＴＴＴＧＣ＾３Ｃ＾０ＡＧＴＡＴＣＴＡ
Ａ＾４Ａ＾０ＧＣＧＧＡＣＴＡＡ＾５Ａ＾０ＧＴＴＧＡ
ＧＣＴＴ＾６Ｔ＾０ＡＧＣＡＴＴＴＡＡ＾７Ｃ＾０ＴＴ
ＴＡＡＡＣＴＴ＾８Ａ＾０ＡＣＡＡＡＡＡＡＣ＾９Ｃ＾
０Ｔ（III） ＧＴＧＡＴＣＴＣＧ＾１Ｔ＾０ＴＡＧＣＣＴＣＧＧ＾２
Ｃ＾０ＴＡＴＡＴＣＴＣＴ＾３Ａ＾０ＴＡＧＴＴＴＴＧ
Ｃ＾４Ｇ＾０ＡＧＡＡＴＧＴＣＡ＾５Ａ＾０Ｇ（IV） ＣＴＴＧＧＴＧＡＡ＾１Ｇ＾０ＣＴＴＧＡＡＧＧＣ＾２
Ｔ＾０ＣＣＴＴＴＧＡＡＴ＾３Ａ＾０（Ｖ） ＣＧＧＡＣＴＡＡＡ＾１Ｇ＾０ＴＴＧＡＧＣＴＴＴ＾２
Ａ＾０ＧＣＡＴＴＴＡＡＣ＾３＾０Ｔ（２）請求項（１
）記載のヌクレオチド塩基配列をその一部として含む組
換えＤＮＡ。 （３）請求項（１）記載のヌクレオチド塩基配列をその
一部として含む合成ＤＮＡ。 （４）請求項（１）、（２）、及び（３）のいずれか１
項記載のヌクレオチド塩基配列を含むＤＮＡをプローブ
として用いて、ＤＮＡハイブリダイゼーション法により
豚流行性肺炎マイコプラズマ（マイコプラズマ・ハイオ
ニュウモニエ）を検出する方法。[Scope of Claims] (1) A DNA having a nucleotide base sequence selected from the group consisting of the following formulas (I) to (V). (I) CCCCATTTC^1T^0TGGTGAAGC^2
T^0TGAAGGCTC^3C^0TTTGAATA
A^4A^0AATTCATGC^5A^0AATTT
TTAT^6C^0A(II) CCGCTAACC^1T^0CTTCATAAT^2
T^0CTATTTTGC^3C^0AGTATCTA
A^4A^0GCGGACTAA^5A^0GTTGA
GCTT^6T^0AGCATTTAA^7C^0TT
TAAAACTT^8A^0ACAAAAAAC^9C^
0T(III) GTGATCTCG^1T^0TAGCCTCGG^2
C^0TATATCTCT^3A^0TAGTTTTG
C^4G^0AGAATGTCA^5A^0G(IV) CTTGGTGAA^1G^0CTTGAAGGC^2
T^0CCTTTGAAT^3A^0 (V) CGGACTAAA^1G^0TTGAGCTTT^2
A^0GCATTTAAC^3^0T (2) Claim (1
) A recombinant DNA containing the nucleotide base sequence described in (a) as a part thereof. (3) A synthetic DNA comprising the nucleotide base sequence according to claim (1) as a part thereof. (4) Any one of claims (1), (2), and (3)
A method for detecting Mycoplasma swine epidemic pneumonia (Mycoplasma hyopneumoniae) by a DNA hybridization method using DNA containing the nucleotide base sequence described in Section 1 as a probe.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1198475A JPH0361487A (en) | 1989-07-31 | 1989-07-31 | Detection of swine epidemic pneumonia mycoplasma |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1198475A JPH0361487A (en) | 1989-07-31 | 1989-07-31 | Detection of swine epidemic pneumonia mycoplasma |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0361487A true JPH0361487A (en) | 1991-03-18 |
Family
ID=16391728
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1198475A Pending JPH0361487A (en) | 1989-07-31 | 1989-07-31 | Detection of swine epidemic pneumonia mycoplasma |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0361487A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1327070C (en) * | 2001-09-18 | 2007-07-18 | 福伸工业株式会社 | Cloth treatment device |
| US7361746B2 (en) | 1999-12-15 | 2008-04-22 | Gen-Probe Incorporated | Methods and compositions for detection of Mycobacterium avium complex species |
-
1989
- 1989-07-31 JP JP1198475A patent/JPH0361487A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7361746B2 (en) | 1999-12-15 | 2008-04-22 | Gen-Probe Incorporated | Methods and compositions for detection of Mycobacterium avium complex species |
| CN1327070C (en) * | 2001-09-18 | 2007-07-18 | 福伸工业株式会社 | Cloth treatment device |
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