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1. WO2020087557 - QUALITATIVE AND QUANTITATIVE DETECTION METHOD FOR SPODOPTERA EXIGUE NUCLEAR POLYHEDROSIS VIRUS AND APPLICATION THEREOF

Publication Number WO/2020/087557
Publication Date 07.05.2020
International Application No. PCT/CN2018/114548
International Filing Date 08.11.2018
IPC
C12Q 1/70 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
70involving virus or bacteriophage
C12Q 1/686 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6844Nucleic acid amplification reactions
686Polymerase chain reaction
G01N 33/569 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
569for microorganisms, e.g. protozoa, bacteria, viruses
CPC
C12Q 1/04
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
02involving viable microorganisms
04Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
C12Q 1/686
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6844Nucleic acid amplification reactions
686Polymerase chain reaction [PCR]
C12Q 1/701
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
70involving virus or bacteriophage
701Specific hybridization probes
C12Q 2600/16
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
2600Oligonucleotides characterized by their use
16Primer sets for multiplex assays
G01N 33/56983
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
569for microorganisms, e.g. protozoa, bacteria, viruses
56983Viruses
Applicants
  • 武汉武大绿洲生物技术有限公司 WUHAN UNIOASIS BIOLOGICAL TECHNOLOGY CO., LTD [CN]/[CN]
Inventors
  • 陈娇 CHEN, Jiao
  • 陶佩文 TAO, Peiwen
  • 尹宜农 YIN, Yinong
  • 林春鸿 LIN, Chunhong
  • 杨未 YANG, Wei
  • 胡家鑫 HU, Jiaxin
  • 杨婷 YANG, Ting
  • 张恒 ZHANG, Heng
  • 刘取 LIU, Qu
  • 贾军 JIA, Jun
  • 彭晖 PENG, Hui
Agents
  • 北京汇泽知识产权代理有限公司 BEIJING HUIZE INTELLECTUAL PROPERTY AGENCY CO., LTD
Priority Data
201811287910.731.10.2018CN
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) QUALITATIVE AND QUANTITATIVE DETECTION METHOD FOR SPODOPTERA EXIGUE NUCLEAR POLYHEDROSIS VIRUS AND APPLICATION THEREOF
(FR) PROCÉDÉ DE DÉTECTION QUALITATIVE ET QUANTITATIVE POUR LE VIRUS DE LA POLYÉDROSE NUCLÉAIRE SPODOPTERA EXIGUA ET SON APPLICATION
(ZH) 甜菜夜蛾核型多角体病毒的定性定量检测方法及应用
Abstract
(EN)
Provided by the present invention is a qualitative and quantitative detection method for Spodoptera exigue nuclear polyhedrosis virus, comprising the following steps: 1) purifying viral particles by employing a combination of sedimentation and differential centrifugation, or a combination of sedimentation, differential centrifugation and density gradient centrifugation; 2) performing qualitative identification on the purified viral particles obtained in step 1) by employing an enzyme-linked immunosorbent assay or PCR-specific primers; 3) preparing the purified viral particles obtained in step 1) into a virus suspension having a certain concentration, and quantitatively measuring the content of the viral particles by employing an electron microscope counting or a microscope counting. The described detection method has the advantages of simple and quick operation, good specificity, high sensitivity, low hardware requirements, and so on; moreover, all types of PCR instruments currently on the market may be used, while the qualitative and quantitative detection of Spodoptera exigue nuclear polyhedrosis virus may be performed quickly, thus providing a reliable method for ensuring product quality for production.
(FR)
La présente invention concerne un procédé de détection qualitative et quantitative pour le virus de la polyédrose nucléaire spodoptera exigua, comprenant les étapes suivantes : 1) purification de particules virales en employant une combinaison de sédimentation et de centrifugation différentielle ou une combinaison de sédimentation, de centrifugation différentielle et de centrifugation par gradient de densité ; 2) réalisation d'une identification qualitative sur les particules virales purifiées obtenues à l'étape 1) en employant un test ELISA ou des amorces spécifiques à la PCR ; 3) préparation des particules virales purifiées obtenues à l'étape 1) dans une suspension de virus ayant une certaine concentration et mesure quantitative du contenu des particules virales à l'aide d'un comptage au microscope électronique ou d'un comptage au microscope. Le procédé de détection décrit présente les avantages d'un fonctionnement simple et rapide, d'une bonne spécificité, d'une sensibilité élevée, d'exigences de matériel faible, etc. De plus, tous les types d'appareils de PCR actuellement sur le marché peuvent être utilisés, permettant d'effectuer rapidement la détection qualitative et quantitative du virus de la polyédrose nucléaire spodoptera exigua, ce qui permet d'obtenir un procédé fiable pour garantir la qualité du produit pour la production.
(ZH)
本发明提供了一种甜菜夜蛾核型多角体病毒的定性定量检测方法,包括如下步骤:1)采用沉降法与差速离心法结合,或沉降法、差速离心法和密度梯度离心法相结合的方法对病毒粒子纯化;2)对步骤1)得到的纯化后的病毒粒子采用酶联免疫法或PCR特异引物法对病毒粒子进行定性鉴别;3)将步骤1)得到的纯化后的病毒粒子制成一定浓度的病毒悬液,采用电子显微镜计数法或显微镜计数法对病毒粒子的含量进行定量测定。该检测方法具有操作简单、快速、特异性好、灵敏度高、对硬件要求低等优势,不仅可使用目前存在市场上的所有类型PCR仪器,而且能快速地对甜菜夜蛾核型多角体病毒进行定性定量检测,为生产提供可靠的保证产品质量的方法。
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