CN102994648B - 一种基于毛细电泳的呼吸道病毒的多重基因检测方法 - Google Patents
一种基于毛细电泳的呼吸道病毒的多重基因检测方法 Download PDFInfo
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- CN102994648B CN102994648B CN201210206565.6A CN201210206565A CN102994648B CN 102994648 B CN102994648 B CN 102994648B CN 201210206565 A CN201210206565 A CN 201210206565A CN 102994648 B CN102994648 B CN 102994648B
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InfluenzaA | 所有甲型流感病毒 |
InfluenzaB | 所有乙型流感病毒 |
GeH1N1 | 所有甲型流感H1N1亚型 |
SeH1N1 | 季节性甲型流感H1N1病毒 |
SWH1N1 | 猪流感H1N1病毒 |
InfAN1_TS | 猪流感病毒达菲敏感 |
InfAN1_TR | 猪流感病毒达菲抗性 |
InfAH3 | 季节性甲型流感病毒H3 |
H5N1 | 禽流感病毒H5N1 |
InfAN1 | 甲型流病毒N1 |
InfAN2 | 甲型流感病毒N2 |
HRV | 人类鼻病毒 |
HCoV-NL63/229E | 人类冠状病毒-NL63/229E |
HCoV-HKU1/OC43 | 人类冠状病毒-HKU1/OC43 |
SARS-CoV | 非典冠状病毒 |
HPIV-1 | 人类副流感病毒1型 |
HPIV-2 | 人类副流感病毒2型 |
HPIV-3 | 人类副流感病毒3型 |
HMPV | 人类偏肺病毒 |
HRSV | 人呼吸道合胞病毒 |
HAdV | 人类腺病毒 |
BocaV | 博卡病毒 |
RNaseP_DNA | 人DNA内参 |
B2M_RNA | 人RNA内参 |
pcDNA3.1 | 反应内参 |
检测内容/内参 | 反转录引物 | PCR引物 |
所有甲型流感病毒 | GGTATGCTTTTGCAMTGARTAGAGG | GCTGCAGTCCTCGCTCACT |
所有乙型流感病毒 | CACACAGGGCTCATAGCAGA | TCTCAGCACTCCAATGTTGC |
所有甲型流感H1N1亚型 | GCMGAYCWRVGAGCACAVARAAT | CCAAAGTTCTTTCATTTTCCART |
季节性甲型流感H1N1病毒 | GGTATGCTTTTGCAMTGARTAGAGG | AAGGGATATTCCTTARTCCTGTAACCAT |
猪流感H1N1病毒 | GCATTCGCAATGGAAAGAAA | TCCTCAATCCTGTGGCCAG |
猪流感病毒达菲敏感 | CGAAATGAATGCCGCTAATCATC | CAGATTCTGGTTGAAAGACACC |
猪流感病毒达菲抗性 | AGTCGAAATGAATGGCCACTAATAATT | CAGATTCTGGTTGAAAGACACC |
季节性甲型流感病毒H3 | CCAAATTGAAGTGACTAATGCTACTGA | TAGTGACCTAAGGGAGGGATAATC |
禽流感病毒H5N1 | CATCAACACTAAACCAGAGATTGG | TCCCCTTTCTTGACAATTTTGTAT |
甲型流感病毒N1 | GGRGCCTTGYTRAATGAACA | ACACATGCACATTCAGAYTCTTG |
甲型流感病毒N2 | CATTGWTTCCAGTTATGTRTGYTC | GTTCTTCCCATCCAYAGRTCA |
人类鼻病毒 | GAAACACGGACACCCAAAGT | GAGTCCTCCGGCCCC |
人类冠状病毒-NL63/229E | CATCATRGTRTATTTAGTAAGATYMTG | CRGTTATGGAMCACGAGCA |
人类冠状病毒-HKU1/OC43 | ATCCCAWTGACAATCCAAAKGG | GAATGTTGCTAAGTAYACTCARTTATG |
非典冠状病毒 | GCAAAACCTGAATCAGTGCC | GTGCTGTGATCATTCGTGGT |
人类副流感病毒1型 | CAATCACAGAATTAATCAGACAAGAA | TGTGTGAGATCTCTGCTTTGC |
人类副流感病毒2型 | AACCATTTACCTAAGTGATGG | GATTGAATCTTTCAATAAAGG |
人类副流感病毒3型 | CTGTGTTCAACTCCMAAAGTTGA | CCTGGTCCAACAGATGGGTA |
人类偏肺病毒 | TGCAGGRRTCMACTGTTTACTAC | AGAGGDGAYAGTGCAACCAT |
人呼吸道合胞病毒 | GGAGCYATWGTGTCATGCTA | TCATAGWAATTTATTATWGGTTC |
人类腺病毒 | CCCAGHGCRTTGTRVACGTA | |
人类腺病毒 | CACTTACCGTCAGTTGAAAACGT | |
博卡病毒 | TCCTGTTGAATGWGTAGGACAAA | |
博卡病毒 | TRCARTGGTTGYTTYAATGAGTAC | |
人DNA内参 | CGCTAGGAATCAGACCAACAC | |
人DNA内参 | ATGGCGGTGTTTGCAGAT | |
人RNA内参 | ATTCATCCAATCCAAATGCG | CCGTGTGAACCATGTGACTT |
反应内参 | CCGAGATAGGGTTGAGTGTTGT | |
反应内参 | AGGGTGATGGTTCACGTAGTG |
步骤 | 温度 | 时间 |
1 | 48℃ | 1分钟 |
2 | 42℃ | 60分钟 |
3 | 95℃ | 5分钟 |
4 | 4℃ | 持续:直至收取RT产物 |
PCR反应试剂 | 量/孔 |
PCR缓冲液,5X | 4μL |
25mM MgCl2 | 4μL |
PCR引物 | 2μL |
DNA聚合酶 | 1.4μL |
RT产物 | 8.6μL |
Total | 20μL |
步骤 | 温度 | 时间 |
1 | 95℃ | 10分钟 |
2 | 94℃ | 30秒钟 |
3 | 60℃ | 30秒钟 |
4 | 70℃ | 1分钟 |
5 | N/A | 重复2-4步骤34次(共35次) |
6 | 70℃ | 1分钟 |
7 | 4℃ | 持续:直至收取PCR产物 |
GeXP样品 | 量/孔 |
SLS上样液 | 38.75μL |
DNA大小标准400 | 0.5μL |
PCR产物 | 1μL |
Total | 40μL |
矿物油 | 1滴 |
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Families Citing this family (4)
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MX2013009755A (es) * | 2013-08-23 | 2015-02-23 | Univ Autonoma De Nuevo Leon | Método para la detección múltiple de virus de influenza tipo a, tipo b, o tipo c mediante oligonucleotidos diseñados para permitir discriminación alelica en una reacción de rt-pcr multiplex. |
CN104342503B (zh) * | 2014-10-29 | 2016-11-30 | 福建国际旅行卫生保健中心 | 一种同时检测12种常见呼吸道病毒的方法 |
CN106048096A (zh) * | 2016-08-16 | 2016-10-26 | 无锡市疾病预防控制中心 | 一种同时检测四种呼吸道病毒的rt‑pcr方法所用的四对上下游引物 |
CN106222304A (zh) * | 2016-08-16 | 2016-12-14 | 无锡市疾病预防控制中心 | 一种同时检测五种呼吸道病毒的rt‑pcr方法所用的五对上下游引物 |
Citations (1)
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CN102618662A (zh) * | 2012-04-26 | 2012-08-01 | 重庆出入境检验检疫局检验检疫技术中心 | 6种食源性致病菌的GeXP多重快速检测用引物和检测方法 |
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Publication number | Priority date | Publication date | Assignee | Title |
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CN102618662A (zh) * | 2012-04-26 | 2012-08-01 | 重庆出入境检验检疫局检验检疫技术中心 | 6种食源性致病菌的GeXP多重快速检测用引物和检测方法 |
Non-Patent Citations (2)
Title |
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GeXP多重PCR技术同时检测12种常见呼吸道病毒;李瑾等;《病毒学报》;20111130;第27卷(第6期);摘要、材料与方法部分 * |
未知病原和病毒性出血热检测方法的研究;孙玉兰;《中国博士学位论文全文数据库 医药卫生科技辑》;20120115(第1期);第114-120页 * |
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