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1. (WO2017099445) GENETIC MARKER FOR DISCRIMINATING AND DETECTING CAUSATIVE BACTERIA OF FISH EDWARDSIELLOSIS AND STREPTOCOCCOSIS, AND METHOD FOR DISCRIMINATING AND DETECTING CAUSATIVE BACTERIA USING SAME
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/2017/099445 International Application No.: PCT/KR2016/014215
Publication Date: 15.06.2017 International Filing Date: 06.12.2016
IPC:
C12Q 1/68 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
68
involving nucleic acids
Applicants:
대한민국(관리부서:국립수산과학원) REPUBLIC OF KOREA(NATIONAL FISHERIES RESEARCH AND DEVELOPMENT INSTITUTE) [KR/KR]; 부산시 기장군 기장읍 기장해안로 216 216, Gijanghaean-ro Gijang-eup Gijang-gun Busan 46083, KR
Inventors:
김명석 KIM, Myoung Sug; KR
정승희 JUNG, Sung-Hee; KR
최혜승 CHOI, Hye Sung; KR
한현자 HAN, Hyun-Ja; KR
도정완 DO, Jeung-Wan; KR
김진도 KIM, Jin-Do; KR
Agent:
이처영 LEE, Cheo Young; KR
장제환 CHANG, Je Hwan; KR
Priority Data:
10-2015-017687011.12.2015KR
Title (EN) GENETIC MARKER FOR DISCRIMINATING AND DETECTING CAUSATIVE BACTERIA OF FISH EDWARDSIELLOSIS AND STREPTOCOCCOSIS, AND METHOD FOR DISCRIMINATING AND DETECTING CAUSATIVE BACTERIA USING SAME
(FR) MARQUEUR GÉNÉTIQUE PERMETTANT DE DISTINGUER ET DE DÉTECTER DES BACTÉRIES À L'ORIGINE DE L'INFECTION À EDWARDSIELLA ET DE LA STREPTOCOCCOCIE CHEZ LE POISSON, ET PROCÉDÉ DE DIFFÉRENCIATION ET DE DÉTECTION DE TELLES BACTÉRIES UTILISANT LEDIT MARQUEUR
(KO) 어류 에드와드 감염증과 연쇄구균 감염증 원인세균의 판별 및 검출용 유전자 마커, 및 이를 이용한 원인세균의 판별 및 검출 방법
Abstract:
(EN) The present invention relates to a genetic marker for discriminating and detecting the causative bacteria of fish edwardsiellosis and streptococcosis in fish, and a method for discriminating and detecting the causative bacteria using the same. More specifically, the present invention relates to a method for selecting and amplifying a genetic marker comprising a bacteria-specific single nucleotide polymorphism (SNP) of a DNA base sequence which encodes 16S rDNA genes of Edwardsiella tarda, streptococcal Streptococcus iniae, Streptococcus parauberis and Lactococcus garvieae, which are the causative bacteria of fish edwardsiellosis and streptococcosis in fish, then hybridizing peptide nucleic acids (PNA) which specifically recognize an amplified product, obtaining a melting curve for each temperature through temperature control of the hybridized product, and discriminating bacteria species or detecting whether a fish was infected with the bacteria species from the melting temperature through an analysis of the obtained melting curve. The present invention has the effects of discriminating the causative bacteria of fish diseases in a simple, rapid, and accurate manner and detecting whether the fish was infected with the bacteria, by selecting a genetic marker for discriminating and/or detecting Edwardsiella tarda, Streptococcus iniae, Streptococcus parauberis and Lactococcus garvieae which are the causative bacteria of fish diseases and allowing bacteria species to represent fluorescence amplification and a melting curve different from each other by using a peptide nucleic acid and primer pair specific to the genetic marker.
(FR) La présente invention concerne un marqueur génétique permettant de distinguer et de détecter les bactéries à l'origine de l'infection à Edwardsiella et de la streptococcocie chez le poisson, et un procédé de différenciation et de détection de telles bactéries utilisant ledit marqueur. L'invention concerne spécifiquement un procédé de sélection et d'amplification d'un marqueur génétique comprenant un polymorphisme de nucléotide simple (SNP) spécifique à une bactérie d'une séquence de bases d'ADN qui code pour des gènes d'ADN ribosomique 16S d'Edwardsiella tarda, Streptococcus iniae streptococcale, Streptococcus parauberis et Lactococcus garvieae, qui sont les bactéries à l'origine de l'infection à Ewardsiella et de la streptococcie chez le poisson, le procédé consistant ensuite à hybrider des acides nucléiques peptidiques (ANP) qui reconnaissent spécifiquement un produit amplifié, obtenir une courbe de fusion pour chaque température par régulation de la température du produit hybridé et distinguer les espèces de bactéries ou détecter si un poisson a été infecté par les espèces bactériennes à partir de la température de fusion, par une analyse de la courbe de fusion obtenue. La présente invention permet de distinguer de façon simple, rapide et précise les bactéries à l'origine de maladies chez le poisson et de détecter si le poisson a été infecté par les bactéries, en sélectionnant un marqueur génétique pour distinguer et/ou détecter Edwardsiella tarda, Streptococcus iniae, Streptococcus parauberis et Lactococcus garvieae, qui sont les bactéries à l'origine de maladies chez le poisson, et permettre à des espèces de bactéries de représenter une amplification de fluorescence et une courbe de fusion différentes les uns des autres au moyen d'un acide nucléique peptidique et d'une paire d'amorces spécifiques au marqueur génétique.
(KO) 본 발명은 어류 에드와드 감염증과 연쇄구균 감염증 원인세균의 판별 및 검출용 유전자 마커, 및 이를 이용한 원인세균의 판별 및 검출 방법에 관한 것으로, 더욱 자세하게는 어류 에드와드 감염증과 연쇄구균 감염증 원인세균인 에드와드시엘라 타르다(Edwardsiella tarda), 연쇄구균인 스트렙토코카스 이니애(Streptococcus iniae), 스트렙토코카스 파라우베리스(Streptococcus parauberis) 및 락토코카스 가비에(Lactococcus garvieae)의 16S rDNA 유전자를 코딩하는 DNA 염기서열 중 세균 특이적 단일염기다형성(Single nucleotide polymorphism, SNP)을 포함하는 유전자 마커를 선정하여 증폭시킨 다음, 증폭산물을 특이적으로 인식하는 펩티드핵산(Peptide Nucleic Acid, PNA)을 혼성화시키고, 혼성화된 산물의 온도 조절을 통해 온도별 융해곡선을 얻고, 상기 얻은 융해곡선의 분석을 통해 융해온도로부터 세균 종을 판별하거나 어류의 상기 세균 종의 감염 여부를 검출하는 방법에 관한 것이다. 본 발명은 어류 질병 원인세균인 에드와드시엘라 타르다, 스트렙토코카스 이니애, 스트렙토코카스 파라우베리스 및 락토코카스 가비에의 판별 및/또는 검출용 유전자 마커를 선정하고, 상기 유전자 마커에 특이적인 펩티드핵산 및 프라이머 쌍을 이용하여 세균 종마다 서로 다른 형광의 증폭 및 융해곡선을 나타내게 함으로써 어류 질병 원인세균을 간단ㆍ신속ㆍ정확하게 판별하고, 어류의 상기 세균의 감염 여부를 검출할 수 있는 효과가 있다.
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Organization (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Korean (KO)
Filing Language: Korean (KO)
Also published as:
US20180127798