CN112143788A - 一种鉴定pah基因同时含两个突变位点位置的试剂盒及方法 - Google Patents
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Abstract
本发明公开了一种鉴定PAH基因同时含两个突变位点位置的试剂盒及方法。所述试剂盒中含有如下引物对:F1:GTGTAAATTACTTACTGTTAATGGAATCAG;R1:CCTCTATTACGTGGCAGAGAGTT;F2:GTGTAAATTACTTACTGTTAATGGAATCAT;R2:CCTCTATTACGTGGCAGAGAGTA;所述两个突变位点为:C.1301C>A、C.1174T>A突变位点。本发明能够鉴定C.1301C>A、C.1174T>A突变位点是在同一条染色体,还是不同的染色体上,工作量小,耗时短,易操作。
Description
技术领域
本发明属于生物医学检测技术领域,具体涉及一种鉴定PAH基因同时含两个突变位点(C.1301C>A、C.1174T>A)位置的试剂盒及方法。
背景技术
苯丙氨酸羟化酶(PAH)在苯丙氨酸代谢途径中起重要的作用,当PAH缺乏或者活性减低可导致苯丙氨酸代谢障碍,从而使苯丙氨酸通过其他代谢途径产生苯丙酮酸等有害物质。这些有害的代谢产物达到异常增高的水平后,积蓄于组织、血浆和脑脊液中,并大量从尿中排出,产生苯丙酮尿症(PKU)。PAH基因突变引起经典型PKU,占所有患者的90%左右,本病遗传方式为常染色体隐性遗传,所以对PKU患者进行PAH基因的筛查,能及早的鉴别,诊断,从而达到早发现、早治疗的效果。
不同的PAH基因的突变对PAH活性及结构影响的程度不同。针对于本案中涉及到的两个突变位点,发生在同一条或者不同的染色体上,对患者病情严重程度的影响是不同的。如果两个突变发生在同一条染色体上,则不致病;两个突变分别发生在不同的染色体上,则致病。为了评估疾病的严重程度,鉴定已知的不同的PAH基因突变分布在同一条还是不同染色体上,对疾病的诊断具有决定性作用。现有的方法是提取DNA,构建载体,进行体外表达实验,需要全套的分子生物学及微生物、细胞培养的实验室,难度极大,且工作量大,耗时2-3个月,普通实验室难以实施。
发明内容
本发明旨在克服现有技术的不足,提供一种鉴定PAH基因同时含两个突变位点位置的试剂盒及方法。
为了达到上述目的,本发明提供的技术方案为:
鉴定PAH基因同时含两个突变位点位置的试剂盒中含有如下引物对:
F1:GTGTAAATTACTTACTGTTAATGGAATCAG(SEQ ID NO.1)
R1:CCTCTATTACGTGGCAGAGAGTT(SEQ ID NO.2)
F2:GTGTAAATTACTTACTGTTAATGGAATCAT(SEQ ID NO.3)
R2:CCTCTATTACGTGGCAGAGAGTA(SEQ ID NO.4);
所述两个突变位点为:C.1301C>A、C.1174T>A突变位点。
优选地,引物对按如下排列组合使用:
①:F1+R1
②:F1+R2
③:F2+R1
④:F2+R2。
优选地,根据C.1301C>A、C.1174T>A突变分布情况,引物对按如下排列组合使用:
①:当待测样本同时含C.1301C>A、C.1174T>A突变但分布在不同染色体上时:
F1+R2或F2+R1;其中,F1和R1是检测野生型的引物;
②当待测样本同时含C.1301C>A、C.1174T>A突变且分布在同一条染色体上时:
F2+R2。
基于上述试剂盒鉴定同时含C.1301C>A、C.1174T>A突变位点的方法是采用所述试剂盒中的引物对,在如下次序和条件下进行扩增检测:
①:94℃,5min;
②:94℃,30s;55℃,30s;72℃,2min;35Cycles;
③:72℃,7min;25℃,∝;
若扩增后的片段经琼脂糖凝胶电泳出现特异性条带,则表明存在C.1301C>A、C.1174T>A突变位点。
本发明根据PAH基因已知位点C.1301C>A、C.1174T>A突变,设计两组引物,第一组:3′端碱基与正常的模板碱基互补,第二组:3′端碱基与突变的模板碱基互补。基于耐热TaqDNA聚合酶缺乏3′→5′外切校正活性的特点,引物3′端的特异碱基分别互补于野生型和突变型等位基因的相对碱基,若此碱基对形成错配,链延伸反应就会因3′,5′-磷酸二酯键形成障碍而受阻,所以其扩增结果为:野生模板阻碍,突变引物放大;或突变模板阻碍,野生引物放大。
本发明设计了不同的引物对,将提取的模板扩增后进行琼脂糖凝胶电泳从而达到鉴定的目的。本发明能够鉴定C.1301C>A、C.1174T>A突变位点是在同一条染色体,还是不同的染色体上。现有的方法是提取DNA、构建克隆载体、构建克隆载体、体外表达实验,需要全套的分子生物学及微生物、细胞培养的实验室,难度极大,且工作量大,耗时2-3个月,普通实验室难以实施,本发明仅需要引物设计、提取DNA、扩增与跑胶,能够在1天内完成所有试验,工作量小,耗时短,易操作成本极低,且普通分子实验室均可操作。
附图说明
图1为Z、T样本电泳图;
图2为S样本电泳图;
图3为对比引物对电泳图。
具体实施方式
1.DNA提取
选取3个样本,一个野生型,命名为Z,一个同时含C.1301C>A、C.1174T>A突变但分布在不同染色体上的样本,命名为T,另一个同时含C.1301C>A、C.1174T>A突变且分布在同一条染色体上的样本,命名为S。根据试剂QIAamp DNA Blood Mini Kit(250)(凯杰,163030523)说明书提取DNA,保存于-20℃备用。
2.PCR扩增
根据表1所示反应体系进行片段扩增(商用试剂购自南京诺唯赞生物科技有限公司)。
表1反应体系
试剂 | 体积(μL) |
2×Vazyme LAmp Master Mix(商用) | 25 |
ddH2O | 13 |
Primers Mixer(F+R) | 8 |
模板DNA(商用) | 4 |
总体积 | 50 |
按照表1配置反应体系,其中,Primers Mixer(F+R)为扩增引物。
本发明设计了两组扩增引物序列,如下表2。其中F1、F2序列3’端是针对位点C.1301C>A野生型和突变型模板,R1、R2序列3’端是针对位点C.1174T>A野生型和突变型模板。表2引物序列按5P浓度1:1F端与R端自由组合,可得4组Primers Mixer(F+R)混合物,如表3。
表2引物序列
表3Primers Mixer(F+R)引物组合
第1组 | F1+R1 |
第2组 | F1+R2 |
第3组 | F2+R1 |
第4组 | F2+R2 |
所有试剂配制完成后,将每组引物分装成3份,分别加入样本Z、T和S,得12份混合液,按照引物分组命名为Z1-Z4,T1-T4和S1-S4,并按照下列扩增程序进行PCR扩增:
[94℃,5min];[94℃,30s;55℃,30s;72℃,2min;35Cycles];[72℃,7min;25℃,∝]。
3.电泳
①配置凝胶
取1g琼脂糖凝胶(干粉)加入100ul 1X TAE溶液混合。将混合液放入微波炉,设置微波炉“大火-3min”。待冷却后加入5ul DD核酸染料,混匀倒胶待凝固。
②点样
用水稀释到1X loading buffer。取出待测扩增产物,按照产物:(loadingbuffer)=1:1比例在配置好的胶块上加样。同时在一泳槽内加入DL 2000DNA Marker。
③电泳
设置好电泳仪参数:70V-100min。
4.结果
结果如图1和图2所示,Z样本在第一组引物组合中出现特异性条带,其余组合未出现条带。T样本在第2组,第3组出现了特异性条带,第1组,第4组未出现条带。S样本在第4组引物组合中出现特异性条带,第1、2、3组均未出现。
5.对照
对照组的引物序列如表4,引物组合如表5,其他操作同具体实施方式的1至3。
表4对照引物序列
表5Primers Mixer(F+R)对照引物组合
第1组 | F1+R1 |
第2组 | F1+R2 |
第3组 | F2+R1 |
第4组 | F2+R2 |
结果如图3所示,Z样本除了在第一组引物组合中出现特异性条带,在不应该出现条带的其余组合也出现了条带;T样本除了在第2组、第3组出现了特异性条带,在不应该出现条带的第1组,第4组也出现了条带;S样本除了在第4组引物组合中出现特异性条带,在不应该出现条带的第1、2、3组也出现了条带。
序列表
<110> 长沙金域医学检验实验室有限公司
<120> 一种鉴定PAH基因同时含两个突变位点位置的试剂盒及方法
<141> 2020-09-10
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Claims (4)
1.一种鉴定PAH基因同时含两个突变位点位置的试剂盒,其特征在于,所述试剂盒中含有如下引物对:
F1:GTGTAAATTACTTACTGTTAATGGAATCAG
R1:CCTCTATTACGTGGCAGAGAGTT
F2:GTGTAAATTACTTACTGTTAATGGAATCAT
R2:CCTCTATTACGTGGCAGAGAGTA;
所述两个突变位点为:C.1301C>A、C.1174T>A突变位点。
2.如权利要求1所述的试剂盒,其特征在于,引物对按如下排列组合使用:
①:F1+R1
②:F1+R2
③:F2+R1
④:F2+R2。
3.如权利要求2所述的试剂盒,其特征在于,根据C.1301C>A、C.1174T>A突变分布情况,引物对按如下排列组合使用:
①当待测样本同时含C.1301C>A、C.1174T>A突变但分布在不同染色体上时:F1+R2或F2+R1;
②当待测样本同时含C.1301C>A、C.1174T>A突变且分布在同一条染色体上时:F2+R2。
4.一种鉴定PAH基因同时含两个突变位点位置的方法,其特征在于,所述方法是采用权利要求1至3任一项所述试剂盒中的引物对,在如下次序和条件下进行扩增检测:
①:94℃,5min;
②:94℃,30s;55℃,30s;72℃,2min;35Cycles;
③:72℃,7min;25℃,∝;
若扩增后的片段经琼脂糖凝胶电泳出现特异性条带,则表明存在C.1301C>A、C.1174T>A突变位点。
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CN103436616A (zh) * | 2013-08-23 | 2013-12-11 | 苏州市立医院 | 中国人群苯丙酮尿症pah基因筛查试剂盒 |
CN105755109A (zh) * | 2015-11-23 | 2016-07-13 | 苏州市立医院 | 一种新的苯丙酮尿症基因筛查和诊断体系及试剂盒 |
-
2020
- 2020-09-10 CN CN202010945008.0A patent/CN112143788A/zh active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103436616A (zh) * | 2013-08-23 | 2013-12-11 | 苏州市立医院 | 中国人群苯丙酮尿症pah基因筛查试剂盒 |
CN105755109A (zh) * | 2015-11-23 | 2016-07-13 | 苏州市立医院 | 一种新的苯丙酮尿症基因筛查和诊断体系及试剂盒 |
Non-Patent Citations (2)
Title |
---|
BARBARA ARBEITHUBER等: "Haplotyping of Heterozygous SNPs in Genomic DNA Using Long-Range PCR", METHODS MOL BIOL, pages 3 - 22 * |
LI N.N.等: "Molecular characterisation of phenylketonuria in a Chinese mainland population using nextgeneration sequencing", SCIENTIFIC REPORTS, pages 15769 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111073884A (zh) * | 2020-02-14 | 2020-04-28 | 昆明理工大学 | 提高非编码区域距离<50bp的SNP中具有功能效应的SNP位点检测准确率的方法 |
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