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1. CN105524986 - LAMP detection method for rapidly detecting Candidatus Liberibacter asiaticus

Office China
Application Number 201511003710.0
Application Date 29.12.2015
Publication Number 105524986
Publication Date 27.04.2016
Grant Number 105524986
Grant Date 18.09.2018
Publication Kind B
IPC
C12Q 1/6844
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
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; 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
02involving viable microorganisms
04Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
C12R 1/01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C-C12Q75
1Microorganisms
01Bacteria or actinomycetales
CPC
C12Q 1/6844
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
Applicants INSPECTION AND QUARANTINE COMPREHENSIVE TECHNOLOGY CENTER, JIANGXI ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU
江西出入境检验检疫局检验检疫综合技术中心
Inventors HUANG LILI
黄丽莉
LI YIN
李茵
ZHU JIANXIN
祝建新
LUO TAOPENG
罗涛朋
LIU YING
刘英
Title
(EN) LAMP detection method for rapidly detecting Candidatus Liberibacter asiaticus
(ZH) 一种快速检测亚洲柑桔黄龙病菌的LAMP检测方法
Abstract
(EN)
The invention discloses an LAMP detection method for rapidly detecting Candidatus Liberibacter asiaticus. The method comprises the following steps: 1, designing primers: designing four LAMP specific primers by adopting 16SrDNA of Candidatus Liberibacter asiaticus as a target gene; 2, extracting nucleic acid: extracting citrus blade total DNA through adopting a CTAB technology; and 3, detecting the LAMP specificity: taking a 5 * Reaction Buffer, F3 and B3 primers, FIP and BIP primers, Mg<2+>, dNTPs, Bst, DNA polymerase and a template, adding ddH2O to a constant volume of 25[mu]L, placing a reaction PCR tube in a constant temperature incubator, reacting, and inactivating to end the reaction. The method has the advantages of simple operation, low requirements of environment and facility and devices, low cost, rapidness and accuracy, is especially suitable for large-scale primary screening of Liberobacter asiaticum, and provides a new way for detecting and controlling Liberobacter asiaticum.

(ZH)
本发明公开了一种快速检测亚洲柑桔黄龙病菌的LAMP检测方法,包括以下步骤:1)引物设计:采用柑桔黄龙病菌的16SrDNA作为靶基因,设计4条LAMP特异性引物;2)核酸提取:按照CTAB法提取柑桔叶片总DNA;3)LAMP特异性检测:取5×Reaction Buffer、F3和B3引物、FIP和BIP引物、Mg2+、dNTPs、Bst DNA聚合酶、模板、加ddH2O定容至25μL;将反应PCR管置于恒温孵育器反应,之后灭活结束反应;本发明操作简便、对环境设施设备要求不高、成本低,快速准确,特别适合对于亚洲柑桔黄龙病的大规模初筛,为柑桔黄龙病的检测与防控提供了一种新的手段。