JP2012523831A - アベリノ角膜ジストロフィー診断用プライマー - Google Patents
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Abstract
Description
さらに、本発明は、配列番号25乃至配列番号42からなる群から選択される塩基配列で表されるアベリノ角膜ジストロフィー診断用リアルタイムPCRプローブを提供する。
BIGH3遺伝子のエクソン4の変異を含む部分を増幅させるプライマーをPrimer Express3.0ソフト(Applied Biosystems U.S.A.)を用いて、配列番号1〜2、配列番号3〜4、配列番号5〜6、配列番号7〜8、配列番号9〜10、配列番号11〜12、配列番号13〜14、配列番号15〜16、配列番号17〜18、配列番号19〜20、配列番号21〜22及び配列番号23〜24のプライマー対を設計した。
ACD Re primer : 5'-CCA TCT CAG GCC TCA GCT T(配列番号2)(60bp)
AV Fw primer : 5'-TGC AGC CCT ACC ACT CTC AA(配列番号3)
AV Re primer : 5'-AGG CCT CGT TGC TAG G(配列番号4)(150bp)
Real Fw primer : 5'-TAG TCT CTT ATT CTA ATA GA(配列番号5)
Real Re primer : 5'-GCT GCA GAC TCT GTG TTT AA(配列番号6)(860bp)
ACD Fw2 primer : 5'-CCA TCC CTC CTT CTG TCT TCT G(配列番号7)
ACD Re2 primer : 5'-CGG GCC CCT CCA TCT C(配列番号8)(140bp)
ACD Fw3 primer : 5'-CAG AGA AGG GAG GGT GTG GTT(配列番号9)
ACD Re3 primer : 5'-GGG CGA AGA TGG TGA AGC T(配列番号10)(190bp)
ACD Fw4 primer : 5'-TCC TCG TCC TCT CCA CCT GTA(配列番号11)
ACD Re4 primer : 5'-AGC TGG CAA GGA GGC CC(配列番号12)
ACD Fw5 primer : 5'-TTT GGG CTT TCC CAC ATG C(配列番号13)
ACD Re5 primer : 5'-GGC AGA CGG AGG TCA TCT CA(配列番号14)
ACD Fw6 primer : 5'-GTA GTA CCG TGC TCT CTG(配列番号15)
ACD Re6 primer : 5'-AGT TCC CCA TAA GAA TCC CCC(配列番号16)
ACD Fw7 primer : 5'-GGC TGG ACC CCC AGA GG(配列番号17)
ACD Re7 primer : 5'-ACC CCT CGG GGA AGT AAG G(配列番号18)
ACD Fw8 primer : 5'-AAC CTT TAC GAG ACC CTG GGA(配列番号19)
ACD Re8 primer : 5'-GAC TCC CAT CCA TCA TGC CC(配列番号20)
ACD Fw9 primer : 5'-AGT CGT TGG ATC CAC CAC CA(配列番号21)
ACD Re9 primer : 5'-GAC GTC ATT TCC TAC TGT TTC AGG(配列番号22)
ACD Fw10 primer : 5'-CCC CCC AGA AAC AGC CTG(配列番号23)
ACD Re10 primer : 5'-TTC TAA GGG GTT AAG GAG AAA GCT T(配列番号24)
変異がない正常遺伝子断片と結合するプローブはVICで標識し、変異がある遺伝子断片と結合するプローブはFAMで標識し、相補的な遺伝子断片との結合が容易になるよう、MGB(minor groove binder)を付けた。
Mutant probe1 : FAM-CAC GGA CCA CAC GGA-NFQ(配列番号26)
Normal probe2 : VIC-ACA CGG ACC GCA CG-NFQ(配列番号27)
Mutant probe2 : FAM-ACA CGG ACC ACA CG-NFQ(配列番号28)(14bp)
Normal probe3 : VIC-TAC ACG GAC CGC A-NFQ(配列番号29)
Mutant probe3 : FAM-TAC ACG GAC CAC A-NFQ(配列番号30)(13bp)
Normal probe4 : VIC-CTG TAC ACG GAC CGC ACG-NFQ(配列番号31)
Mutant probe4 : FAM-CTG TAC ACG GAC CAC ACG-NFQ(配列番号32)(18bp)
Normal probe5 : VIC-CTG TAC ACG GAC CGC ACG GAG-NFQ(配列番号33)
Mutant probe5 : FAM-CTG TAC ACG GAC CAC ACG GAG-NFQ(配列番号34)(21bp)
Normal probe6 : VIC-GCT GTA CAC GGA CCG CAC GGA GAA-NFQ(配列番号35)
Mutant probe6 : FAM-GCT GTA CAC GGA CCA CAC GGA GAA-NFQ(配列番号36)
Normal probe7 : VIC-ACC GCA CGG AGA AGC-NFQ(配列番号37)
Mutant probe7 : FAM-ACC ACA CGG AGA AGC-NFQ(配列番号38)
Normal probe8 : VIC-ACC GCA CGG AGA AGC TGA GGC-NFQ(配列番号39)
Mutant probe8 : FAM-ACC ACA CGG AGA AGC TGA GGC-NFQ(配列番号40)
Normal probe8 : VIC-ACC GCA CGG AGA AGC TGA GGC CTG-NFQ(配列番号41)
Mutant probe8 : FAM-ACC ACA CGG AGA AGC TGA GGC CTG-NFQ(配列番号42)
検査対象者の血液、毛根、口腔上皮細胞からサンプルを採取し、DNAを分離した。DNA分離精製には、フェノール/クロロホルム抽出法(Miller, SA et al., Nucl. Acids Res. 16:1215, 1988)を一部改変して実施し、分離されたDNAは、TE緩衝液(10mM Tris−Cl,1mM EDTA,pH7.4)適量に溶解させた後、1%アガローズゲルで電気泳動して確認して、鋳型DNAとして用いた。
Claims (3)
- 配列番号1〜2、配列番号3〜4、配列番号5〜6、配列番号7〜8、配列番号9〜10、配列番号11〜12、配列番号13〜14、配列番号15〜16、配列番号17〜18、配列番号19〜20、配列番号21〜22及び配列番号23〜24からなる群から選択される塩基配列で表されるアベリノ角膜ジストロフィー診断用リアルタイムPCRプライマー対。
- 配列番号25〜配列番号42からなる群から選択される塩基配列で表されるアベリノ角膜ジストロフィー診断用リアルタイムPCRプローブ。
- VICまたはFAMで標識されている請求項2に記載のアベリノ角膜ジストロフィー診断用リアルタイムPCRプローブ。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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KR10-2009-0033528 | 2009-04-17 | ||
KR1020090033528A KR101251538B1 (ko) | 2009-04-17 | 2009-04-17 | 아벨리노 각막이상증 진단용 프라이머 |
PCT/KR2009/007099 WO2010120027A1 (ko) | 2009-04-17 | 2009-12-01 | 아벨리노 각막이상증 진단용 프라이머 |
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JP2012523831A true JP2012523831A (ja) | 2012-10-11 |
JP5738842B2 JP5738842B2 (ja) | 2015-06-24 |
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JP2012505796A Active JP5738842B2 (ja) | 2009-04-17 | 2009-12-01 | アベリノ角膜ジストロフィー診断用プライマー |
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US (4) | US20120077200A1 (ja) |
EP (2) | EP2848689A1 (ja) |
JP (1) | JP5738842B2 (ja) |
KR (1) | KR101251538B1 (ja) |
CN (3) | CN104774930A (ja) |
AU (1) | AU2009344501A1 (ja) |
BR (1) | BRPI0924016A2 (ja) |
CA (1) | CA2759222A1 (ja) |
IL (1) | IL215845A0 (ja) |
RU (1) | RU2534817C2 (ja) |
SG (1) | SG175732A1 (ja) |
WO (1) | WO2010120027A1 (ja) |
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Cited By (2)
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JP2017501685A (ja) * | 2013-11-15 | 2017-01-19 | アベリノ ラボ ユーエスエー インコーポレイテッドAvellino Lab USA, Inc. | 眼疾患に関連したアレルのマルチプレックス検出方法 |
US11987809B2 (en) | 2015-11-13 | 2024-05-21 | Avellino Lab Usa, Inc. | Methods for the treatment of corneal dystrophies |
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KR101125212B1 (ko) * | 2010-10-01 | 2012-03-21 | (주)아벨리노 | 아벨리노 각막이상증 진단용 시스템 |
KR101480522B1 (ko) | 2013-02-15 | 2015-01-08 | 솔젠트 (주) | 아벨리노 각막 이상증 판별용 진단 키트 |
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KR101577109B1 (ko) * | 2013-04-23 | 2015-12-11 | 주식회사 녹십자엠에스 | 아벨리노 각막이상증 진단용 조성물 및 이의 진단방법 |
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KR101125212B1 (ko) | 2010-10-01 | 2012-03-21 | (주)아벨리노 | 아벨리노 각막이상증 진단용 시스템 |
EP3068908B1 (en) | 2013-11-15 | 2021-01-06 | Avellino Lab USA, Inc. | Methods for multiplex detection of alleles associated with ophthalmic conditions |
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2009
- 2009-04-17 KR KR1020090033528A patent/KR101251538B1/ko active IP Right Grant
- 2009-12-01 RU RU2011146553/10A patent/RU2534817C2/ru active
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- 2009-12-01 WO PCT/KR2009/007099 patent/WO2010120027A1/ko active Application Filing
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2017501685A (ja) * | 2013-11-15 | 2017-01-19 | アベリノ ラボ ユーエスエー インコーポレイテッドAvellino Lab USA, Inc. | 眼疾患に関連したアレルのマルチプレックス検出方法 |
JP2020039339A (ja) * | 2013-11-15 | 2020-03-19 | アベリノ ラボ ユーエスエー インコーポレイテッドAvellino Lab USA, Inc. | 眼疾患に関連したアレルのマルチプレックス検出方法 |
JP7077289B2 (ja) | 2013-11-15 | 2022-05-30 | アベリノ ラボ ユーエスエー インコーポレイテッド | 眼疾患に関連したアレルのマルチプレックス検出方法 |
US11525160B2 (en) | 2013-11-15 | 2022-12-13 | Avellino Lab Usa, Inc. | Methods for multiplex detection of alleles associated with ophthalmic conditions |
US11987809B2 (en) | 2015-11-13 | 2024-05-21 | Avellino Lab Usa, Inc. | Methods for the treatment of corneal dystrophies |
Also Published As
Publication number | Publication date |
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SG175732A1 (en) | 2011-12-29 |
US11268146B2 (en) | 2022-03-08 |
WO2010120027A1 (ko) | 2010-10-21 |
CA2759222A1 (en) | 2010-10-21 |
EP2420574A1 (en) | 2012-02-22 |
CN102459593B (zh) | 2015-04-08 |
KR101251538B1 (ko) | 2013-04-08 |
US20150093751A1 (en) | 2015-04-02 |
US20180274034A1 (en) | 2018-09-27 |
IL215845A0 (en) | 2012-01-31 |
EP2848689A1 (en) | 2015-03-18 |
BRPI0924016A2 (pt) | 2016-10-11 |
CN104774930A (zh) | 2015-07-15 |
US20120077200A1 (en) | 2012-03-29 |
ZA201107967B (en) | 2013-08-28 |
RU2011146553A (ru) | 2013-05-27 |
CN102459593A (zh) | 2012-05-16 |
US20220205044A1 (en) | 2022-06-30 |
JP5738842B2 (ja) | 2015-06-24 |
CN110872613A (zh) | 2020-03-10 |
KR20100115030A (ko) | 2010-10-27 |
US9938581B2 (en) | 2018-04-10 |
EP2420574A4 (en) | 2012-08-29 |
RU2534817C2 (ru) | 2014-12-10 |
EP2420574B1 (en) | 2014-10-29 |
AU2009344501A1 (en) | 2011-11-17 |
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