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WO2006000140A1 - Amorces, sequences sondes et procedes utiles dans plusieurs dosages rt-pcr fluorescents en temps reel des sous-types h5, h7 et h9 du virus de la grippe aviaire - Google Patents

Amorces, sequences sondes et procedes utiles dans plusieurs dosages rt-pcr fluorescents en temps reel des sous-types h5, h7 et h9 du virus de la grippe aviaire Download PDF

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Publication number
WO2006000140A1
WO2006000140A1 PCT/CN2005/000667 CN2005000667W WO2006000140A1 WO 2006000140 A1 WO2006000140 A1 WO 2006000140A1 CN 2005000667 W CN2005000667 W CN 2005000667W WO 2006000140 A1 WO2006000140 A1 WO 2006000140A1
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WIPO (PCT)
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sequence
primer
avian influenza
probe
bases
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PCT/CN2005/000667
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English (en)
Chinese (zh)
Inventor
Jingzhong Lin
Xinglong Xiao
Shengli Liu
Anqing Zhong
Zhifeng Qin
Jianqian Lv
Xianzhong Jin
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Taitai Genomics, Inc.
Shenzhen Entry-Exitinspection And Quarantine Bureau Of The People's Republic Of China
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Publication of WO2006000140A1 publication Critical patent/WO2006000140A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes

Definitions

  • the present invention relates to RT-PCR amplification primers and probes for highly pathogenic avian influenza H5, H7 and H9 subtype nucleotide fragments, and simultaneous typing assays for avian influenza ⁇ 5, ⁇ 7 and ⁇ 9 subtypes .
  • Avian Influenza also known as European chicken or true chicken, is an acute highly contagious disease caused by influenza A virus. It is an internationally recognized devastating disease that can pass pigs or Other animals are transmitted to humans and therefore have important ecological and public health implications, especially for highly pathogenic avian influenza, which is classified as a Class A infectious disease by the World Organisation for Animal Health. '
  • avian flu not only seriously harms birds, but also harms mammals, especially those that can infect people and cause death.
  • the avian influenza virus belongs to the RNA virus. Like all RNA viruses, due to the lack of corrective function of RNA polymerase, the genome has a high error rate when it is transcribed, and its genome is segmented, which is prone to rearrangement. Virus has high
  • the amino acid sequence of the HA cleavage site determines the virulence of avian influenza virus.
  • the HA cleavage site of highly pathogenic avian influenza has 6 consecutive sites.
  • Basic amino acids, while low pathogenic avian influenza has 2 basic amino acids.
  • RT-PCR reverse transcription polymerase chain reaction
  • RNA is reverse transcribed into cDNA and then the DNA fragment is rapidly amplified in vitro using DNA polymerase.
  • Fluorescence real-time RT-PCR is based on common RT-PCR.
  • a pair of primers is added to the amplification reaction system, and a specific fluorescent probe is added.
  • the probe is labeled with a fluorescent reporter group at both ends.
  • a fluorescent quenching group of oligonucleotides When the probe is intact, the fluorescent signal emitted by the reporter group is absorbed by the quenching group, so that the fluorescent signal emitted by the fluorescent group of the probe is not detected, and at the 5' end of the Taq enzyme during PCR amplification.
  • Exonuclease activity cleaves the fluorescent probe that specifically binds to the amplified fragment of interest, so that the fluorescent reporter group is released from the reaction system, and the shielding effect of the fluorescence quenching group is removed. At this time, the fluorescence report can be detected.
  • the fluorescence signal of the group, the amount of fluorescence signal is proportional to the amount of amplification product.
  • Multiple real-time fluorescent PCR is a method of adding multiple pairs of primers and probes in the same reaction system and simultaneously detecting multiple target fragments, which is of great significance in the identification of disease-like diseases and the typing of 'multi-serotype pathogens.
  • the fluorescent labeling probe is used to further improve the specificity and accuracy of the detection;
  • the detection is more sensitive by automatically detecting the change of the fluorescent signal; Perform a fully enclosed detection reaction without post-processing the PCR product to avoid contamination; d. Monitor the results in real time,
  • the object of the present invention is to provide primers, probe sequences and methods for multiple real-time fluorescent RT-PCR detection of avian influenza H5, H7 and H9 subtypes.
  • the object of the present invention can be achieved by the following technical solutions: Primers and fluorescent probes are designed separately by analyzing all reported avian influenza H5, H7 and H9 subtype genomic sequences. Based on the conventional RT-PCR detection technique, a nucleotide probe labeled with two fluorophores is added, and the fluorescent reporter group (R) is labeled at the 5' end of the probe, and the fluorescence quenching group (Q) Marked at the 3' end of the probe, the two constitute an energy transfer structure, that is, the fluorescence emitted by the fluorescent reporter group can be absorbed by the fluorescence quenching group, and when the distance is far, the inhibition is weakened, and the reporter group is weakened. The fluorescence signal is enhanced.
  • the probe hybridizes to the amplified fragment of interest on the template. Since the Taq enzyme has the exonuclease activity at the 5' end to the 3' end, the probe is cleaved during the amplification extension phase, and the fluorescence quenching group is The inhibition of Q) disappeared, and the fluorescent signal of the reporter group was enhanced to detect the H5, H7 and H9 subtypes of avian influenza.
  • the principle of fluorescence RT-PCR detection is shown in Figure 1. Primer and probe design: Multiple pairs of primers and probes were designed by comparing the genomic sequences of all reported avian influenza virus H5, H7 and H9 subtypes, respectively, and selecting non-secondary structure and highly conserved nucleotide segments.
  • the primers are generally about 20 bases in length, and there are no complementary sequences between the primers and the primers, and they should specifically target the avian influenza virus H5, H7 and H9 subtypes, and there is no cross reaction between them, which can be in one reaction tube. It is also used to distinguish between H5, H7 and H9 subtypes.
  • the primer sequences and probe sequences for nucleotide amplification for detecting the avian influenza H5 subtype include: the upstream primer ⁇ 5 ⁇ 673 sequence is GACCAGCTACCATGATTGCCA and the complementary sequence is CTGGTCGATGGTACTAACGGT, and the downstream primer H5pr l 600 sequence is GGAGTCAAATTTGGA ATCAATGG and the complementary sequence is The primer pair consisting of CCTCAGTTTAA ACCTTAGTTACC and the upstream primer position of the primer pair extend 10 bases toward the 5' end, and the downstream primer position extends 10 bases toward the 3' end and 10 bases toward the 5' end.
  • the primer sequence and the complementary sequence obtained in the region range is TGCTAGGGAACTCGCCA, the complementary sequence is ACGATCCCTTGAGCGGT, and the 3 ⁇ 4 probe extends 10 bases toward the 3' end and 10 base regions toward the 5' end. Probe sequences and complementary sequences obtained in the range.
  • Primer sequences and probe sequences for nucleotide amplification for detecting avian influenza H7 subtype include -
  • the primer pair consisting of the upstream primer H7pfl292 sequence is CGTGTCCAATTAATGACATTTCC and the complementary sequence is GCACAGGTTAATTACTGTAAAGG
  • the downstream primer H7prl202 sequence is ATCACAGGCAA ATTGAATCGT and the complementary sequence is TAGTGTCCGTTTAACTTAGCA
  • the upstream primer position of the primer pair extends 10 bases toward the 5' end.
  • the probe H7pbl247 sequence is TTTGAGCTGATAGACAATGA and the complementary sequence is AAACTCGACTATCTGTTACT, and
  • the probe extends 10 bases in the 3' end direction and a probe sequence and a complementary sequence extending in the range of 10 bases in the 5' end direction.
  • the primer pair consisting of the upstream primer ⁇ 7 ⁇ 664 sequence is GCCATTGCAATGGGCCT and the complementary sequence is CGPTA ACGTTACCCGGA
  • the downstream primer H7prl736 sequence is AGTAGAAAC 'AAGGGTGTTTTTTCCA and the complementary sequence is TCATCTTTGTTCCC ACAAAAAAGGT
  • the upstream primer position of the primer pair extends toward the 5' end.
  • the primer sequence and the complementary sequence obtained by extending the base primer position by 10 bases in the 3' end direction and extending 10 bases in the 5' end direction;
  • the probe H7pbl712 sequence is CGGTGCACTATTTGTATA and the complementary sequence is GCCACGTGATAAACATAT
  • Primer sequences and probe sequences for nucleotide amplification for detecting avian influenza H9 subtype include: CCTGTGACACATGCCAAAGA from the upstream primer H9pfl51 sequence and GGACACTGTGT ACGGTTTCT as the complementary sequence, CTTTCGACRATGTAGGACCATTC for the downstream primer H9pr302 sequence and GAAAGCTGYTACATCCTGGTAAG for the complementary sequence
  • the primer pair and the upstream primer position of the primer pair extend 10 bases in the 5' end direction, and the downstream primer position extends 10 bases in the 3' end direction and 10 bases in the 5' end direction.
  • the primer sequence and the complementary sequence; the probe H9pbl75 sequence is CTCCACACAGAGCACAAT and the complementary sequence is GAGGT GTGTCTCGTGTTA, and the probe is extended to 10 bases in the 3' end direction and 10 bases in the 5' end direction. Needle sequence and complementary sequence.
  • the salient features of the present invention are: Fully utilizing the high-efficiency amplification of PCR technology, the good specificity of nucleic acid hybridization, and the rapid sensitivity of fluorescence detection technology, a high-pathogenic avian influenza H5 and a single detection operation can be performed on one sample.
  • the detection of H7 and H9 subtypes can determine the serotype, and has the advantages of reliable and accurate results, simple operation, time and labor saving, reduced detection cost and improved detection efficiency.
  • BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 Schematic diagram of real-time fluorescence RT-PCR detection of Taqman probe.
  • FIG. 1 Avian influenza H5, H7 and H9 subtypes of triple real-time fluorescence RT-PCR detection curve
  • Figure 3 Avian influenza H5 and H9 subtype double real-time fluorescence RT-PCR detection curve.
  • Figure 4 Avian influenza H7 and H9 subtypes of double real-time fluorescence RT-PCR detection curve.
  • Figure 5 Avian influenza H5 and H7 subtypes of double real-time fluorescence RT-PCR detection curve.
  • the specific method of establishment and optimization of the reaction system using the inactivated avian influenza virus H5, H7 and H9 subtype strains as samples to be tested, using the QIAamp Viral RNA Mini kit to extract viral genomic RNA, separately stored and stored in a 20 ⁇ spare.
  • the concentration of MgCl 2 is increased from 1 mmol/L to 10 mmol/L in 1 mmol/L under the same conditions in the reaction system, and 5 mmol is selected after repeated experiments.
  • /L is the magnesium ion concentration in the kit reaction system.
  • AMV RnaseXL reverse transcriptase
  • Taq enzyme Optimization of the amount of Taq DNA polymerase (Taq enzyme) By comparing the optimization results of the amount of Taq enzyme (in units of Unit), 5U was selected as the amount of Taq enzyme in the kit reaction system.
  • the probe concentrations of avian influenza H5, H7 and H9 subtypes were from ⁇ . ⁇ ⁇ mol/L to 0.5 ⁇ mol/'L, respectively, under the same conditions in the reaction system. The ratio was measured after serial dilution, and the final concentration of the optimal probe was determined to be 0.2 ⁇ mol/L by analysis and comparison of the test results.
  • Table 1 Avian influenza ⁇ 5, ⁇ 7 and ⁇ 9 subtypes of triple real-time fluorescence RT-PCR reaction of various components
  • each reagent should be adjusted according to the ratio.
  • the collection of the fluorescent signal of the reaction tube in the instrument should be set.
  • the selected fluorescent detection channel is identical to the fluorescent reporter group labeled by the probe. .
  • the specific setting method varies depending on the instrument. Refer to the instrument manual.
  • Method 1 Use QI Aamp Viral RNA Mini kit to extract genomic RNA of avian influenza virus from various sources. The specific steps are as follows:
  • a Take 560 ⁇ l of AVL buffer containing carrier RNA into a 1.5 ml microcentrifuge tube; b. Add 140 ⁇ l of plasma, serum, urine sample, cell culture supernatant or swab to PBS wash solution. Oscillating for 15 sec; - c incubation at room temperature (15-25 ⁇ ) for 10 min;
  • Solubility without RNase (fully boil and mix, 'or repeatedly blow with a gun), store at -20 °C for later use.
  • test results can be monitored in real time.
  • double-phase real-time fluorescent RT-PCR was performed using specific primers and probes for avian influenza ⁇ 5 and ⁇ 9 subtypes to detect positive samples containing genomic RNA of avian influenza virus H5 and H9 subtypes, and the results were obtained.
  • Specific fluorescence curve Figure 3
  • double-phase real-time fluorescent RT-PCR was performed using specific primers and probes for the influenza ⁇ 7 and ⁇ 9 subtypes to detect positive samples containing genomic RNA of avian influenza virus H7 and H9 subtypes.
  • Specific fluorescence curve ( Figure 4).
  • the reagents and positive control nucleotides required for real-time fluorescent RT-PCR were assembled into avian influenza H5, H7 and H9 subtypes.

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Abstract

L'invention se rapporte à des amorces, des séquences sondes et des procédés utiles dans plusieurs dosages RT-PCR fluorescents en temps réel des sous types H5, H7 et H9 du virus de la grippe aviaire. Elle concerne plus précisément des amorces et des sondes pour l'amplification RT-PCR des fragments nucléotidiques des sous-types H5, H7 et H9 de la grippe aviaire et des séquences supplémentaires de ceux-ci, ainsi que des dosages d'éloignement simultané par type des sous-types H5, H7 et H9 du virus de la grippe aviaire.
PCT/CN2005/000667 2004-06-25 2005-05-13 Amorces, sequences sondes et procedes utiles dans plusieurs dosages rt-pcr fluorescents en temps reel des sous-types h5, h7 et h9 du virus de la grippe aviaire WO2006000140A1 (fr)

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CN 200410027815 CN1670221A (zh) 2004-06-25 2004-06-25 用于禽流感h5、h7和h9亚型多重实时荧光rt－pcr检测的引物、探针序列和方法
CN200410027815.5 2004-06-25

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2021516A1 (fr) * 2006-04-28 2009-02-11 Siemens Healthcare Diagnostics Inc. Amorces et sondes d'acide nucléique permettant de détecter les virus de la grippe humaine et de la grippe aviaire
CN102433395A (zh) * 2011-12-27 2012-05-02 中国检验检疫科学研究院 一种检测禽流感病毒通用亚型的液相芯片方法
CN102433396A (zh) * 2011-12-27 2012-05-02 中国检验检疫科学研究院 一种检测禽流感病毒h7亚型的液相芯片方法
CN103276109A (zh) * 2013-05-10 2013-09-04 浙江省疾病预防控制中心 禽流感h7n9病毒rt-pcr检测试剂盒及检测方法
CN111235321A (zh) * 2020-04-15 2020-06-05 福建省农业科学院畜牧兽医研究所 一种番鸭呼肠孤病毒和新型鸭呼肠孤病毒双重TaqMan荧光定量RT-PCR检测试剂盒
CN111518954A (zh) * 2020-05-07 2020-08-11 广西壮族自治区兽医研究所 H5和n8亚型禽流感病毒双重纳米pcr检测的引物组、试剂盒及方法

Families Citing this family (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100368560C (zh) * 2005-09-22 2008-02-13 广州华银医药科技有限公司 一种呼吸道病原体的实时荧光聚合酶链式反应检测方法
CN100378230C (zh) * 2005-12-01 2008-04-02 上海交通大学 H5、h7、h9亚型禽流感病毒的实时荧光定量pcr检测方法
CN101058833B (zh) * 2007-02-08 2012-05-23 中国检验检疫科学研究院动植物检疫研究所 用于禽流感病毒检测和分型的引物系统和方法
CN101736090B (zh) * 2008-11-07 2012-06-27 中山大学达安基因股份有限公司 一种实时荧光rt-pcr检测h5亚型禽流感病毒的试剂盒
CN101886136B (zh) * 2009-05-12 2012-10-17 中山大学达安基因股份有限公司 一种实时荧光rt-pcr检测甲型h1n1流感病毒的试剂盒
CN101798602B (zh) * 2010-03-18 2012-04-25 山东出入境检验检疫局检验检疫技术中心 复合检测禽流感和新城疫病毒的试剂盒及检验方法
CN102260749B (zh) * 2011-04-25 2012-12-19 吉林农业大学 H5、h7、h9亚型禽流感病毒检测试剂盒
CN103320528B (zh) * 2013-06-03 2014-12-10 深圳太太基因工程有限公司 一种用于以荧光rt-pcr检测样品中禽流感病毒的引物对、探针和包含其的试剂盒
CN103805719B (zh) * 2014-02-20 2016-03-09 广西壮族自治区兽医研究所 鸡新城疫病毒、h9亚型禽流感病毒和禽肺炎病毒三重rt-pcr试剂盒及其应用
CN103789454B (zh) * 2014-02-26 2016-05-25 广西壮族自治区兽医研究所 H9亚型禽流感病毒和鸭坦布苏病毒二重rt-pcr检测试剂盒
CN103993105B (zh) * 2014-06-13 2016-01-27 广西壮族自治区兽医研究所 鸭坦布苏病毒和禽流感病毒多重pcr检测试剂盒
CN104531901B (zh) * 2014-12-31 2017-08-25 四川圣迪乐村生态食品股份有限公司 一种同时检测禽4种呼吸道疾病病原的pcr检测方法
CN106435024B (zh) * 2016-09-26 2020-02-14 南京农业大学 检测禽流感病毒亚型的荧光定量pcr引物、探针和试剂盒及检测方法
CN107937611B (zh) * 2017-12-18 2020-09-04 北京卓诚惠生生物科技股份有限公司 检测禽流感病毒亚型h5、h7和h9的引物探针组
CN107937622A (zh) * 2018-01-16 2018-04-20 北京大有泰莱生物技术有限公司 一种禽流感病毒检测方法及其试剂盒
CN108300809A (zh) * 2018-02-23 2018-07-20 湖南国测生物科技有限公司 一种禽流感h5、h7以及h9亚型的荧光pcr检测试剂盒、制备方法及使用方法
CN108950081A (zh) * 2018-08-10 2018-12-07 中国动物卫生与流行病学中心 一种h7亚型禽流感病毒的实时荧光定量rt-pcr检测方法
CN110885901B (zh) * 2018-09-10 2022-11-25 中国动物疫病预防控制中心（农业部屠宰技术中心） 用于鉴别h5和h7高致病变异型禽流感病毒的冻干微芯片、试剂盒及方法

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
LIU Y. ET AL, JOURNAL OF JILIN AGRICULTURAL UNIVERSITY, vol. 26, no. 1, February 2004 (2004-02-01), pages 93 - 96 *
ZHANG H.X. ET AL, INSPECTION AND QUARANTINE SCIENCE, vol. 14, no. 1, February 2004 (2004-02-01), pages 1 - 5 *
ZHU W.S. ET AL, CHINESE JOURNAL OF MEDICINAL GUIDE, vol. 5, no. 1, 2003, pages 27 - 30 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2021516A1 (fr) * 2006-04-28 2009-02-11 Siemens Healthcare Diagnostics Inc. Amorces et sondes d'acide nucléique permettant de détecter les virus de la grippe humaine et de la grippe aviaire
EP2021516A4 (fr) * 2006-04-28 2009-05-20 Siemens Healthcare Diagnostics Amorces et sondes d'acide nucléique permettant de détecter les virus de la grippe humaine et de la grippe aviaire
US7993838B2 (en) 2006-04-28 2011-08-09 Siemens Healthcare Diagnostics Inc. Nucleic acid primers and probes for detecting human and avian influenza viruses
CN101460632B (zh) * 2006-04-28 2012-11-14 西门子医疗保健诊断公司 用于检测人和禽类流感病毒的核酸引物及探针
CN102433395A (zh) * 2011-12-27 2012-05-02 中国检验检疫科学研究院 一种检测禽流感病毒通用亚型的液相芯片方法
CN102433396A (zh) * 2011-12-27 2012-05-02 中国检验检疫科学研究院 一种检测禽流感病毒h7亚型的液相芯片方法
CN102433395B (zh) * 2011-12-27 2013-10-09 中国检验检疫科学研究院 一种检测禽流感病毒通用亚型的液相芯片方法
CN103276109A (zh) * 2013-05-10 2013-09-04 浙江省疾病预防控制中心 禽流感h7n9病毒rt-pcr检测试剂盒及检测方法
CN111235321A (zh) * 2020-04-15 2020-06-05 福建省农业科学院畜牧兽医研究所 一种番鸭呼肠孤病毒和新型鸭呼肠孤病毒双重TaqMan荧光定量RT-PCR检测试剂盒
CN111518954A (zh) * 2020-05-07 2020-08-11 广西壮族自治区兽医研究所 H5和n8亚型禽流感病毒双重纳米pcr检测的引物组、试剂盒及方法

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