CN106435030B - 一种联合检测禽流感h7n9和h9n2的四通道荧光定量pcr检测方法及试剂盒 - Google Patents
一种联合检测禽流感h7n9和h9n2的四通道荧光定量pcr检测方法及试剂盒 Download PDFInfo
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- CN106435030B CN106435030B CN201610991488.8A CN201610991488A CN106435030B CN 106435030 B CN106435030 B CN 106435030B CN 201610991488 A CN201610991488 A CN 201610991488A CN 106435030 B CN106435030 B CN 106435030B
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- gene
- avian influenza
- quantitative pcr
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- 238000001514 detection method Methods 0.000 title claims abstract description 60
- 241000342557 H7N9 subtype Species 0.000 title claims abstract description 44
- 238000003753 real-time PCR Methods 0.000 title claims abstract description 41
- 241001473386 H9N2 subtype Species 0.000 title claims abstract description 39
- 206010064097 Avian influenza Diseases 0.000 title claims abstract description 33
- 208000002979 Influenza in Birds Diseases 0.000 title claims abstract description 33
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 15
- 238000003745 diagnosis Methods 0.000 claims abstract description 5
- 238000003908 quality control method Methods 0.000 claims description 17
- 239000000523 sample Substances 0.000 claims description 14
- 229920001850 Nucleic acid sequence Polymers 0.000 claims description 10
- 229920000160 (ribonucleotides)n+m Polymers 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 241000700605 Viruses Species 0.000 claims description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerine Chemical compound 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OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
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[Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 5
- 101700011961 DPOM Proteins 0.000 claims description 4
- 101710029649 MDV043 Proteins 0.000 claims description 4
- 101700061424 POLB Proteins 0.000 claims description 4
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CCOC(=O)OC(=O)OCC FFYPMLJYZAEMQB-UHFFFAOYSA-N 0.000 description 1
- 230000036826 Excretion Effects 0.000 description 1
- 210000003608 Feces Anatomy 0.000 description 1
- 241000713869 Moloney murine leukemia virus Species 0.000 description 1
- 210000003462 Veins Anatomy 0.000 description 1
- 102000004965 antibodies Human genes 0.000 description 1
- 108090001123 antibodies Proteins 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 201000009910 diseases by infectious agent Diseases 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000002934 lysing Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 230000001954 sterilising Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
本发明公开了一种联合检测禽流感H7N9和H9N2的四通道荧光定量PCR检测方法及其试剂盒。本发明方法能够同时检测出待测标本中禽流感病毒H7N9的H7基因和N9基因，以及禽流感病毒H9N2的H9基因和N2基因。由于该方法引入了特异性扩增引物和荧光探针，使得检测的灵敏度和特异性得到很大程度的增强，从而避免了其他检测方法特异性不高容易漏诊和误诊的问题。同时，该试剂盒根据本方法原理制得的试剂盒，可方便地同时检测出禽流感病毒H7N9的H7基因和N9基因，以及禽流感病毒H9N2的H9基因和N2基因，准确率高。
Description
技术领域
本发明属于分子检测技术领域，具体涉及一种联合检测禽流感H7N9和H9N2的四通道荧光定量PCR检测方法及试剂盒。
背景技术
在我国新发的人感染H7N9亚型病毒引起了广泛的关注，病毒的常监测、早诊断、预警成为极其重要工作。研究表明，禽源H9N2亚型禽流感病毒为H7N9亚型流感这一新型重组病毒提供了内部基因。然而，鸡源H9N2亚型禽流感病毒的流行和进化是如何促进新型禽流感H7N9出现的机制尚不清楚，中国农业大学刘金华教授团队对该机制进行了研究，研究表明在过去长达10多年中，多种H9N2基因型共同存在，G57基因型出现并随着抗体特性变化更加适应鸡宿主。在2010～2013年期间，也就是新型禽流感病毒H7N9出现前期，H9N2亚型禽流感病毒G57基因型成为免疫鸡农场的主要基因型，并引起了鸡场禽流感的广泛爆发，最终为新型禽流感H7N9提供了内部基因。禽农场中H9N2病毒的流行和变化监测可以为可能造成流感大流行的新型禽流感病毒出现提供重要的监测和早期诊断预警。
随着分子生物学的快速发展，多重PCR已广泛应用于禽病混合感染的鉴别诊断，其原理是设计能够同时扩增出多种待测病原特定片段大小的多对引物，在PCR反应体系中加入多对引物，能够对多种病原进行鉴别诊断。多重RT-PCR因其灵敏性、特异性和操作的简单性被广泛应用于禽病的多重检测，但结果判定需要电泳，费时费力，且反应产物容易产生污染而导致假阳性，而多重PCR是靠片段的大小来区别的，一般到了三重以上，片段大小差别大，其每种病毒的扩增效率不一样，造成结果存在偏差。荧光定量PCR技术融合了PCR的多种优点，通过直接检测PCR反应过程中荧光信号的变化实现对目的分子的定量，不需要电泳检测，并且整个过程完全闭管式操作，污染机率降低，避免了常规PCR容易造成的假阳性问题。相对常规PCR，荧光定量PCR在敏感性、特异性与速度等方面具有优势，但实时荧光PCR技术受到荧光种类及仪器自身的限制，最多只能对5个靶标进行检测，且实验成功的难度极大。
发明内容
本发明的一个目的在于提供一种用于联合检测禽流感H7N9和H9N2的荧光定量PCR引物组。
本发明的另一个目的在于提供一种用于联合检测禽流感H7N9和H9N2的荧光定量PCR试剂盒。
本发明的另一个目的在于提供一种用于联合检测禽流感H7N9和H9N2的荧光定量PCR检测方法。
本发明所采取的技术方案是：
一种用于联合检测禽流感H7N9和H9N2的荧光定量PCR引物组，其包括用于检测禽流感H7N9亚型的H7基因的引物对及荧光探针、用于检测禽流感H7N9亚型的N9基因的引物对及荧光探针、用于检测禽流感H9N2亚型的H9基因的引物对及荧光探针、用于检测禽流感H9N2亚型的N2基因的引物对及荧光探针，用于检测H7、N9、H9、N2基因的荧光探针报告基团不相同。
作为优选的，所述用于检测禽流感H7N9亚型的H7基因的引物对及荧光探针的核苷酸序列如下所示：
H7-F：5’-TGCACTGCATGTTTCCATTCTT-3’(SEQ ID NO.1)；
H7-R：5’-GGCTACAAAGATGTGATACTTTGGTTTAG-3’(SEQ ID NO.2)；
H7-P：5’FAM-ATGAAAACAAGGCCCATTGCAATGGC-BHQ13’(SEQ ID NO.3)。
作为优选的，所述用于检测禽流感H7N9亚型的N9基因的引物对及荧光探针的核苷酸序列如下所示：
N9-F：5’-TCGCGCCCTGATAAGCT-3’(SEQ ID NO.4)；
N9-R：5’-GCATTCCACCCTGCTGTTG-3’(SEQ ID NO.5)；
N9-P：5’VIC-CCACTATCATCACCGCCCACAGTGT-BHQ13”(SEQ ID NO.6)。
作为优选的，所述用于检测禽流感H9N2亚型的H9基因的引物对及荧光探针的核苷酸序列如下所示：
H9-F：5’-CAATGGGGTTTGCTGCCT-3’(SEQ ID NO.7)；
H9-R：5’-TTATATACAAATGTTGCATCTGC-3’(SEQ ID NO.8)；
H9-P：5’TexasRed-TTCTGGGCCATGTCCAATGGTTCT-BHQ23’(SEQ ID NO.9)。
作为优选的，所述用于检测禽流感H9N2亚型的N2基因的引物对及荧光探针的核苷酸序列如下所示：
N2-F：5’-TGACACTGCACTTCAAGCAA-3’(SEQ ID NO.10)；
N2-R：5’-TTGGTTCACATGTCACTACTTGAT-3’(SEQ ID NO.11)；
N2-P：5’CY5-ATGAATGCAGCATCCCCTCGAAC-BHQ23’(SEQ ID NO.12)。
一种用于联合检测禽流感H7N9和H9N2的荧光定量PCR试剂盒，其包含上述的荧光定量PCR引物组。
所述试剂盒还包含病毒RNA提取液、Tth DNA聚合酶、四通道荧光定量PCR Mix、阳性质控品、阴性质控品，所述四通道荧光定量PCR Mix中含有2×四通道荧光定量PCRbuffer、dNTPs。
所述2×四通道荧光定量PCR buffer中含有以下成分：Tricine pH8.7、Mg(OAc)2、KOAc、甘油、DMSO、N,N,N-三甲基甘氨酸。
一种联合检测禽流感H7N9和H9N2的四通道荧光定量PCR检测方法，包括如下步骤：
1)提取待测样品中的病毒RNA；
2)以得到的总RNA为模板，使用上述的荧光定量PCR引物组进行四通道荧光定量PCR
检测；
3)反应结束后，根据待测样品的荧光Ct值判断，待测样品是否为禽流感病毒H7基因、
N9基因、H9基因、N2基因阳性。
四通道荧光定量PCR反应条件为：
95℃1分钟；58℃20分钟；95℃1分钟；
95℃10秒，58℃70秒，68℃10s，采集荧光信号，40个循环。
本发明的有益效果是：
本发明方法能够同时检测出待测标本中禽流感病毒H7N9的H7基因和N9基因，以及禽流感病毒H9N2的H9基因和N2基因。由于该方法引入了特异性扩增引物和荧光探针，使得检测的灵敏度和特异性得到很大程度的增强，从而避免了其他检测方法特异性不高容易漏诊和误诊的问题。同时，该试剂盒根据本方法原理制得的试剂盒，可方便地同时检测出禽流感病毒H7N9的H7基因和N9基因，以及禽流感病毒H9N2的H9基因和N2基因，准确率高。
该方法使用Tth DNA聚合酶，代替传统的Taq酶和MMLV酶的组合，在更高的温度下进行逆转录，提升了四通道扩增这种复杂模版的扩增效果。因此该试剂盒具有广泛的应用前景，适合在各级动物卫生监督机构、疾病控制单位等大规模筛查的推广应用。
附图说明
图1为四通道荧光PCR体系中禽流感病毒H7基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，H7基因检测的灵敏度为1×102cp/μl。
图2为四通道荧光PCR体系中禽流感病毒N9基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，N9基因检测的灵敏度为1×102cp/μl。
图3为四通道荧光PCR体系中禽流感病毒H9基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，H9基因检测的灵敏度为1×102cp/μl。
图4为四通道荧光PCR体系中禽流感病毒N2基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，N2基因检测的灵敏度为1×102cp/μl。
图5为四通道荧光PCR体系中禽流感病毒H7基因检测的特异性实验结果，检测哈兽研H7抗原、H7N9棉拭子标本、H7N7棉拭子标本为阳性，检测哈兽研H5N1抗原、H5N1疫苗、H9N2棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
图6为四通道荧光PCR体系中禽流感病毒N9基因检测的特异性实验结果，检测H7N9棉拭子标本为阳性，检测H7N7棉拭子标本、哈兽研H5N1抗原、华农H5N1疫苗、H9N2棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
图7为四通道荧光PCR体系中禽流感病毒H9基因检测的特异性实验结果，检测H9N2棉拭子标本为阳性，检测H7N7棉拭子标本、哈兽研H5N1抗原、华农H5N1疫苗、H7N9棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
图8为四通道荧光PCR体系中禽流感病毒N2基因检测的特异性实验结果，检测H9N2棉拭子标本为阳性，检测H7N7棉拭子标本、哈兽研H5N1抗原、华农H5N1疫苗、H7N9棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
具体实施方式
下面结合实施例对本发明做进一步的说明，但并不局限于此。
实施例1
1、特异性引物及探针的设计
根据禽流感病毒(Avian influenza virus)H1～H16的核酸序列，以及N1～N9的核酸序列，设计用于检测禽流感病毒H7N9、H9N2基因的引物和探针(在life technology公司合成)。其序列如下：
2、标本采集和预处理
本方法及试剂盒适用标本类型包括禽类的组织、血清、咽拭子、分泌排泄物等。组织标本：无菌条件下取组织约100～200mg左右，置入1.5ml洁净EP管中，保存待检。血清：用一次性无菌注射器抽取受检鸟静脉血3-5ml，留取血清，保存待检。咽及泄殖腔等拭子：用棉拭子取咽或泄殖腔分泌物，将拭子放入盛有1.0ml PBS的离心管中，立即送检。分泌排泄物：无菌条件下采集，保存待检。采集的标本应该尽快送检，或者保存于-20℃。
3、标本RNA提取
3.1RNA提取液的组分：
裂解液
洗涤液
3.2使用杭州莱枫生物生产的RNA纯化柱进行纯化，以如下步骤提取：
1)200μl标本加入500μl裂解液，室温静止1min；
2)12000rpm离心5分钟，弃套管中液体；
3)加入600μL洗涤液，振荡混匀；
4)12000rpm离心5分钟，弃套管中液体；
5)加入600μL洗涤液，振荡混匀；
6)12000rpm离心5分钟，弃套管，换新的1.5mL离心管；
7)加入50μL DEPC处理水，振荡混匀；
8)12000rpm离心5分钟，管中液体即为所提取的RNA。
4、阳性质控品的制备
H7N9假病毒阳性标准品序列如SEQ ID NO.13所示；H9N2假病毒阳性标准品序列如SEQ ID NO.14所示。
将二者分别提交厦门致善生物，合成载体、用MS2噬菌体包装成假病毒颗粒并测定浓度。
返回的H7N9假病毒与H9N2假病毒，以1：1混合，作为试剂盒的阳性质控品，-20±5℃条件下储存。所得假病毒颗粒扩增前需进行RNA提取实验。
5’-GCGGTTATAAAGATGTGATACTTTGGTTTAGCTTCGGGGCATCATGTTTCATACTTCTTGCCATTGCAATGGGCCTTGTCTTCATATGTGTGAAGAATGGAAACATGCGGTGCACTATTTGTATATAATCGCGCCCTGATAAGCTGGCCACTATCATCACCGCCCACAGTATACAACAGCAGGGTGGAATGCATTGGG-3’(SEQ ID NO.13)
5’-CAATGGGGTTTGCTGCCTTCTTGTTCTGGGCCATGTCCAATGGATCATGCAGGTGCAACATTTGTATATAATGACACTGCACTTCAAGCAAAATGAATGCAGCATCCCCTCGAACAATCAAGTAGTGACATGTGAACCAA-3’(SEQ ID NO.14)
5、阴性质控品的制备
取纯化水，高压灭菌后，贮存于冷藏冰箱(2-8℃)，超过7天未分装的转至-20±5℃条件下储存。
6、四通道荧光定量PCR扩增
荧光定量PCR反应的反应体系总体积为25μl，组成如下：多通道荧光定量PCR Mix20μl、DNA聚合酶1μl，以及提取标本RNA(或者阳性质控品、阴性质控品)4μl；其中四通道荧光定量PCR Mix中含有四通道荧光定量PCR buffer、dNTPs等成分。
2×四通道荧光定量PCR buffer中含有以下成分：
四通道荧光定量PCR Mix组成为：
四通道荧光定量PCR反应条件为：
95℃1分钟；58℃20分钟；95℃1分钟；
95℃10秒，58℃70秒，68℃10s，采集荧光信号，40个循环。
7、结果分析和判定
7.1结果分析条件设定
1)设置基线(baseline)：根据机型不同，设置3～15循环，6～15循环，特殊情况可对基线适当调整。
2)设置阈值(threshold):以阈值线刚超过阴性对照扩增曲线(无规则的噪音线)的最高点。
7.2结果判断如下表：
实施例2灵敏度实验
图1为四通道荧光PCR体系中禽流感病毒H7基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，H7基因检测的灵敏度为1×102cp/μl。
图2为四通道荧光PCR体系中禽流感病毒N9基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，N9基因检测的灵敏度为1×102cp/μl。
图3为四通道荧光PCR体系中禽流感病毒H9基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，H9基因检测的灵敏度为1×102cp/μl。
图4为四通道荧光PCR体系中禽流感病毒N2基因检测的灵敏度实验，从左到右依次为1×106、1×105、1×104、1×103、1×102、1×101cp/μl的标准品的扩增结果。由图可知，N2基因检测的灵敏度为1×102cp/μl。
实施例3特异性实验
图5为四通道荧光PCR体系中禽流感病毒H7基因检测的特异性实验结果，检测哈兽研H7抗原、H7N9棉拭子标本、H7N7棉拭子标本为阳性，检测哈兽研H5N1抗原、H5N1疫苗、H9N2棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
图6为四通道荧光PCR体系中禽流感病毒N9基因检测的特异性实验结果，检测H7N9棉拭子标本为阳性，检测H7N7棉拭子标本、哈兽研H5N1抗原、华农H5N1疫苗、H9N2棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
图7为四通道荧光PCR体系中禽流感病毒H9基因检测的特异性实验结果，检测H9N2棉拭子标本为阳性，检测H7N7棉拭子标本、哈兽研H5N1抗原、华农H5N1疫苗、H7N9棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
图8为四通道荧光PCR体系中禽流感病毒N2基因检测的特异性实验结果，检测H9N2棉拭子标本为阳性，检测H7N7棉拭子标本、哈兽研H5N1抗原、华农H5N1疫苗、H7N9棉拭子标本、广州永顺NDV疫苗及阴性质控品均为阴性。
以上实验结果表明：本发明方法及试剂盒可方便地同时检测出禽流感病毒H7N9的H7基因和N9基因，以及禽流感病毒H9N2的H9基因和N2基因，灵敏度和特异性得到很大程度的增强。
SEQUENCE LISTING
<110> 广东省农业科学院动物卫生研究所
<120> 一种联合检测禽流感H7N9和H9N2的四通道荧光定量PCR检测方法及试剂盒
<130>
<160> 14
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> DNA
<213> 人工序列
<400> 1
tgcactgcat gtttccattc tt 22
<210> 2
<211> 29
<212> DNA
<213> 人工序列
<400> 2
ggctacaaag atgtgatact ttggtttag 29
<210> 3
<211> 26
<212> DNA
<213> 人工序列
<400> 3
atgaaaacaa ggcccattgc aatggc 26
<210> 4
<211> 17
<212> DNA
<213> 人工序列
<400> 4
tcgcgccctg ataagct 17
<210> 5
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<212> DNA
<213> 人工序列
<400> 5
gcattccacc ctgctgttg 19
<210> 6
<211> 25
<212> DNA
<213> 人工序列
<400> 6
ccactatcat caccgcccac agtgt 25
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<211> 18
<212> DNA
<213> 人工序列
<400> 7
caatggggtt tgctgcct 18
<210> 8
<211> 23
<212> DNA
<213> 人工序列
<400> 8
ttatatacaa atgttgcatc tgc 23
<210> 9
<211> 24
<212> DNA
<213> 人工序列
<400> 9
ttctgggcca tgtccaatgg ttct 24
<210> 10
<211> 20
<212> DNA
<213> 人工序列
<400> 10
tgacactgca cttcaagcaa 20
<210> 11
<211> 24
<212> DNA
<213> 人工序列
<400> 11
ttggttcaca tgtcactact tgat 24
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<211> 23
<212> DNA
<213> 人工序列
<400> 12
atgaatgcag catcccctcg aac 23
<210> 13
<211> 198
<212> DNA
<213> H7N9
<400> 13
gcggttataa agatgtgata ctttggttta gcttcggggc atcatgtttc atacttcttg 60
ccattgcaat gggccttgtc ttcatatgtg tgaagaatgg aaacatgcgg tgcactattt 120
gtatataatc gcgccctgat aagctggcca ctatcatcac cgcccacagt atacaacagc 180
agggtggaat gcattggg 198
<210> 14
<211> 140
<212> DNA
<213> H7N9
<400> 14
caatggggtt tgctgccttc ttgttctggg ccatgtccaa tggatcatgc aggtgcaaca 60
tttgtatata atgacactgc acttcaagca aaatgaatgc agcatcccct cgaacaatca 120
agtagtgaca tgtgaaccaa 140
Claims (6)
1.一种用于联合检测禽流感H7N9和H9N2的荧光定量PCR引物组，其包括用于检测禽流感H7N9亚型的H7基因的引物对及荧光探针、用于检测禽流感H7N9亚型的N9基因的引物对及荧光探针、用于检测禽流感H9N2亚型的H9基因的引物对及荧光探针、用于检测禽流感H9N2亚型的N2基因的引物对及荧光探针，用于检测H7、N9、H9、N2基因的荧光探针报告基团不相同；
所述用于检测禽流感H7N9亚型的H7基因的引物对及荧光探针的核苷 酸序列如下所示：
H7-F：5'-TGCACTGCATGTTTCCATTCTT-3'(SEQ ID NO.1)；
H7-R：5'-GGCTACAAAGATGTGATACTTTGGTTTAG-3'(SEQ ID NO.2)；
H7-P：5'-ATGAAAACAAGGCCCATTGCAATGGC-3'(SEQ ID NO.3)；
所述用于检测禽流感H7N9亚型的N9基因的引物对及荧光探针的核苷 酸序列如下所示：
N9-F：5'-TCGCGCCCTGATAAGCT-3'(SEQ ID NO.4)；
N9-R：5'-GCATTCCACCCTGCTGTTG-3'(SEQ ID NO.5)；
N9-P：5'-CCACTATCATCACCGCCCACAGTGT-3'(SEQ ID NO.6)；
所述用于检测禽流感H9N2亚型的H9基因的引物对及荧光探针的核苷 酸序列如下所示：
H9-F：5'-CAATGGGGTTTGCTGCCT-3'(SEQ ID NO.7)；
H9-R：5'-TTATATACAAATGTTGCATCTGC-3'(SEQ ID NO.8)；
H9-P：5'-TTCTGGGCCATGTCCAATGGTTCT-3'(SEQ ID NO.9)；
所述用于检测禽流感H9N2亚型的N2基因的引物对及荧光探针的核苷酸序列如下所示：
N2-F：5'-TGACACTGCACTTCAAGCAA-3'(SEQ ID NO.10)；
N2-R：5'-TTGGTTCACATGTCACTACTTGAT-3'(SEQ ID NO.l1)；
N2-P：5'-ATGAATGCAGCATCCCCTCGAAC-3'(SEQ ID NO.12)。
2.一种用于联合检测禽流感H7N9和H9N2的荧光定量PCR试剂盒，其包含权利要求1所述的荧光定量PCR引物组。
3.根据权利要求2所述的荧光定量PCR试剂盒，其特征在于，所述试剂盒还包含病毒RNA提取液、Tth DNA聚合酶、四通道荧光定量PCR Mix、阳性质控品、阴性质控品，所述四通道荧光定量PCR Mix中含有2×四通道荧光定量PCR buffer，dNTPs。
4.根据权利要求3所述的荧光定量PCR试剂盒，其特征在于，所述2×四通道荧光定量PCR buffer中含有以下成分：Tricine pH8.7、Mg(OAc)2、KOAc、甘油、DMSO、N，N，N-三甲基甘氨酸。
5.一种联合检测禽流感H7N9和H9N2的四通道荧光定量PCR检测方法，包括如下步骤：
1)提取待测样品中的病毒RNA；
2)以得到的总RNA为模板，使用权利要求1所述的荧光定量PCR引物组进行四通道荧光定量PCR检测；
3)反应结束后，根据待测样品的荧光Ct值判断，待测样品是否为禽流感病毒H7基因、N9基因、H9基因、N2基因阳性；
上述方法用于非疾病的诊断与治疗。
6.根据权利要求5所述的四通道荧光定量PCR检测方法，其特征在于，四通道荧光定量PCR反应条件为：
95℃1分钟；58℃20分钟；95℃1分钟；
95℃10秒，58℃70秒，68℃10s，采集荧光信号，40个循环。
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