CN105176995B - Thalassemia α can be detected simultaneously based on hydrolysis probes method21.9The kit that deletion form and α 2 make a variation - Google Patents
Thalassemia α can be detected simultaneously based on hydrolysis probes method21.9The kit that deletion form and α 2 make a variation Download PDFInfo
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- CN105176995B CN105176995B CN201510700873.8A CN201510700873A CN105176995B CN 105176995 B CN105176995 B CN 105176995B CN 201510700873 A CN201510700873 A CN 201510700873A CN 105176995 B CN105176995 B CN 105176995B
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Abstract
Thalassemia α can be detected simultaneously based on hydrolysis probes method the invention discloses a kind of21.9The kit that deletion form and α 2 make a variation.The variations of α 2 are the mutant gene type of the genes of α 2 in alpha globin gene cluster (NG_000006.1), and the genotype is described as HBA2:c.301‑24delinsCTCGGCCC.Mentioned reagent box includes that α can be expanded a pair21.9The primer of the characteristic sequence of allele, the primer of a pair of characteristic sequences that can expand the change heteroalleles of α 2, a pair of primers that can expand reference gene GAPDH fragments；Specific detection α21.9The probe of amplified production, the probe of the variation amplified allele products of specific detection α 2 and the probe of specific detection GAPDH segment amplified productions.Mentioned reagent box can the reaction of single tube single be done directly α 2 and make a variation and α21.9The detection of genotype.
Description
Technical field
The present invention relates to technical field of medical detection, and in particular to a kind of can detect simultaneously in ground based on hydrolysis probes method
Extra large anaemia-α21.9The kit that deletion form and α 2 make a variation.
Background technology
Thalassemia is also referred to as globin dyssynthesis anaemia, referred to as poor.Thalassemia is that Guangxi province is most normal
One of monogenic inheritance disease seen, it is poor with β-ground that the poor species in common ground has α-ground poor.In Chinese population, the poor gene in α-ground
Type is common for deletion form --SEA、-α3.7With-α4.2.2013, healthcare hospital for women ＆ children of Qinzhou City, which detected one kind and be named as, ' admired
The poor deletion form in state type α ground ' rarely poor genotype, because the Molecular Biology Mechanism of the genotype is alpha globin gene cluster
(NG_000006.1) missing of one section of 21.9kb DNA sequence dna in, therefore it is abbreviated as-α21.9(Long J,Yan S,Lao K,
Pang W,Ye X,Sun L.The diagnosis and molecular analysis of a novel 21.9kb
deletion(Qinzhou type deletion)causingα+thalassemia.Blood Cells Mol Dis.2014；
52(4):225-9.).In addition, in the research process of applicant, it was found that a kind of new α-globin gene cluster (NG_
000006.1) the mutant gene type of the genes of α 2, the genotype are described as HBA2 in:c.301-24delinsCTCGGCCC.Shen
Ask someone it has also been found that the genotype has certain distribution probability in Guangxi, to be detected both genotype, it is necessary to establish one
Kind can simultaneously and quick detection HBA2:C.301-24delinsCTCGGCCC allele-α21.9The kit of allele.
The content of the invention
The technical problem to be solved in the present invention is to provide it is a kind of based on hydrolysis probes method can detect simultaneously thalassemia-
α21.9The kit that deletion form and α 2 make a variation.The kit can realize that the reaction of single tube single is done directly HBA2:c.301-
24delinsCTCGGCCC and-α21.9The detection of allele, and sensitivity, stability and accuracy with height, and
Higher specificity.
It is of the present invention that thalassemia-α can be detected simultaneously based on hydrolysis probes method21.9What deletion form and α 2 made a variation
Kit, including amplimer and fluorescence probe, wherein：
The α 2 makes a variation as the mutant gene type of the genes of α 2 in α-globin gene cluster (NG_000006.1), the genotype
It is described as HBA2:C.301-24delinsCTCGGCCC, sequence is specially：
gggttgcgggaggtgtagcgcaggcggcggctgcgggcctgggccCTCGGCCCcactgaccctcttctctgcacagc
tcctaagccactgcctgctggtgaccctggccgcccacctccccgccgagttcacccctgcggtgcacgcctccctg
gacaagttcctggcttctgtgagcaccgtgctgacctccaaataccgttaagctggagcctcggtagccgttcctcc
tgcccgctgggcctcccaacgggccctcctcccctccttgcaccggcccttcctggtctttgaataaagtctgagtg
ggcagcagcctgtgtgtgcctgggttctctctatcccggaatgtgccaacaatggaggtgtttacctgtctcagacc
aagga；
Described amplimer is：- α in α-globin gene cluster can be expanded a pair21.9The characteristic sequence of allele
Primer 2 1.9-F and 21.9-R, HBA2 in α-globin gene cluster can be expanded a pair:c.301-24delinsCTCGGCCC
The primer A2V-F and A2V-R of the characteristic sequence of allele, and a pair of primers that can expand reference gene GAPDH fragments
GAPDH-F and GAPDH-R；
Described fluorescence probe is：The fluorescence probe 21.9- of one specific detection 21.9-F and 21.9-R amplified production
Prob, the fluorescence probe A2V-Prob of specific detection A2V-F and A2V-R an amplified production, a specific detection
The fluorescence probe GAPDH-Prob of GAPDH-F and GAPDH-R amplified productions；
Wherein：
It is described to expand-α in α-globin gene cluster21.9In the primer pair of the characteristic sequence of allele,
21.9-F：5’-GGAGCTTTTCCTTCCCTGGAACG-3’；
21.9-R：5’-TGTGGTTGGAGAATGGAGGTGG-3’；
It is described to expand HBA2 in α-globin gene cluster：C.301-24delinsCTCGGCCC the feature of allele
In the primer pair of sequence,
A2V-F：5’-GGGTTGCGGGAGGTGTAG-3’；
A2V-R：5’-TCCTTGGTCTGAGACAGGTA-3’；
In the primer pair that reference gene GAPDH fragments can be expanded,
GAPDH-F：5’-CCCCACACACATGCACTTACC-3’；
GAPDH-R：5’-CCTAGTCCCAGGGCTTTGATT-3’；
The fluorescence probe 21.9-Prob of the specific detection 21.9-F and 21.9-R amplified productions is：
21.9-Prob：5’-6-FAM-CTGGGGAAGGGTGGGTGGGAATAACAGC-BHQ-1-3’；
The fluorescence probe A2V-Prob of described specific detection A2V-F and A2V-R amplified production is：
A2V-Prob：5’-6-ROX-CCTCGGCCCCACTGACCCTC-BHQ-1-3’；
The fluorescence probe GAPDH-Prob of described specific detection GAPDH-F and GAPDH-R amplified production is：
GAPDH-Prob：5’-CY5-AAAGAGCTAGGAAGGACAGGCAACTTGGC-BHQ-2-3’.
In fluorescence probe of the present invention, FAM refers to Fluoresceincarboxylic acid, and ROX refers to carboxy-X-rhodamine, and CY5 refers to cyanine
Dye molecule 5, BHQ-1 and BHQ-2 refer to fluorescent quenching group.
Conventional and necessary component in kit of the present invention, in addition to some available reagent boxes, such as buffer solution, enzyme
Liquid, MgCl2And dNTP.Specifically, enzyme liquid is Taq polymerase system, including the thermal starting enzyme system available for hydrolysis probes method
Deng；The buffer solution is conventional PCR buffer solutions.When enzyme liquid selection uses health by the GoldStar Taq that produce in century
During DNAPolymerase, buffer solution is preferably then the buffer solution supporting with GoldStar Taq DNAPolymerase.
Using mentioned reagent box quick detection HBA2：And-α c.301-24delinsCTCGGCCC21.9The side of allele
Method, comprise the following steps：
1) sample genomic dna is extracted, prepares DNA profiling；
2) reaction system is prepared, is specially：
By primer 2 1.9-F, 21.9-R, A2V-F, A2V-R, GAPDH-F and GAPDH-R, fluorescence probe 21.9-Prob,
A2V-Prob and GAPDH-Prob；And PCR buffer solutions, enzyme liquid, MgCl2, dNTP, water and DNA profiling are configured to reaction system；
3) pattern detection：The reaction system of preparation is entered into performing PCR amplification respectively, records the fluorescence in each PCR reaction tubes
Signal reaches the cycle-index Cq (size of Cq values can reflect institute's detection template number number) undergone during the thresholding of setting；
4) data analysis and result judgement：The analysis software sentence read result carried according to real-time fluorescence quantitative PCR instrument,
When CY5 passages have Cq value readings, then the sample is normally expanded；
On the premise of CY5 passages have reading, if ROX passages have reading, then it represents that the sample carries HBA2:c.301-
24delinsCTCGGCCC allele；
On the premise of CY5 passages have reading, if ROX passages do not have Cq value readings, then it represents that the sample does not carry HBA2:
C.301-24delinsCTCGGCCC allele；
On the premise of CY5 passages have reading, if FAM passages have Cq value readings, then it represents that the sample carrying-α21.9Equipotential
Gene；
On the premise of CY5 passages have reading, if FAM passages do not have Cq value readings, then it represents that the sample does not carry-α21.9
Allele.
In the step 1) of the above method, DNA profiling is prepared using existing conventional method.
In the step 2) of the above method, the concentration of each component is preferably in reaction system：DNA to be detected：1~3ng/ μ L；
Each primer：0.3~0.5 μm of ol/L；Each fluorescence probe concentration is 0.3~0.5 μm of ol/L；DNA profiling：20~200ng；Magnesium from
Son：1.5~1.9mmol/L；The final volume of reaction system is preferably 20 μ L.
In the step 3) of the above method, PCR amplification conditions are：95 DEG C of pre-degenerations 8~12 minutes, then 95 DEG C 15~30
Second, 60 DEG C are annealed 50~70 seconds, 39~49 circulations, in 60 DEG C of annealing steps end collection fluorescence signals.
Compared with prior art, the method have the characteristics that：
1st, kit of the present invention can realize that the reaction of single tube single is done directly HBA2:c.301-
24delinsCTCGGCCC and-α21.9The detection of allele, and sensitivity, stability and accuracy with height, and
Higher specificity.
2nd, operated without open pipe, reduce the possibility of laboratory PCR primer pollution to the utmost；
3rd, it is fewer than the time required to existing other method using the experimental program of hydrolysis probes, it is also not high to equipment requirement.
Brief description of the drawings
Fig. 1 is the amplification curve of 1# samples in the embodiment of the present invention 1；
Fig. 2 is the amplification curve of 2# samples in the embodiment of the present invention 1；
Fig. 3 is the amplification curve of 3# samples in the embodiment of the present invention 1.
Embodiment
With reference to specific embodiment, the present invention is described in further detail, to more fully understand present disclosure, but
The present invention is not limited to following examples.
Embodiment 1：Using testing result of the kit of the present invention in known pattern sheet
1st, the composition of kit：
(1)-α in α-globin gene cluster can be expanded21.9The primer pair 21.9-F of the characteristic sequence of allele and
21.9-R：
21.9-F：5’-GGAGCTTTTCCTTCCCTGGAACG-3’(SEQ ID NO：1)；
21.9-R：5’-TGTGGTTGGAGAATGGAGGTGG-3’(SEQ ID NO：2)；
(2) HBA2 in α-globin gene cluster can be expanded:C.301-24delinsCTCGGCCC allele is (described
HBA2:C.301-24delinsCTCGGCCC the sequence of allele is specially：
gggttgcgggaggtgtagcgcaggcggcggctgcgggcctgggccCTCGGCCCcactgaccctcttctctgcacagc
tcctaagccactgcctgctggtgaccctggccgcccacctccccgccgagttcacccctgcggtgcacgcctccctg
gacaagttcctggcttctgtgagcaccgtgctgacctccaaataccgttaagctggagcctcggtagccgttcctcc
tgcccgctgggcctcccaacgggccctcctcccctccttgcaccggcccttcctggtctttgaataaagtctgagtg
ggcagcagcctgtgtgtgcctgggttctctctatcccggaatgtgccaacaatggaggtgtttacctgtctcagacc
aagga(SEQ ID NO：3) the primer pair A2V-F and A2V-R of characteristic sequence)：
A2V-F:5’-GGGTTGCGGGAGGTGTAG-3’(SEQ ID NO：4)；
A2V-R:5’-TCCTTGGTCTGAGACAGGTA-3’(SEQ ID NO：5)；
(3) GAPDH-F and GAPDH-R in the primer pair of reference gene GAPDH fragments can be expanded：
GAPDH-F:5’-CCCCACACACATGCACTTACC-3’(SEQ ID NO：6)；
GAPDH-R:5’-CCTAGTCCCAGGGCTTTGATT-3’(SEQ ID NO：7)；
(4) the fluorescence probe 21.9-Prob of specific detection 21.9-F and 21.9-R amplified productions is：
21.9-Prob:5’-6-FAM-CTGGGGAAGGGTGGGTGGGAATAACAGC-BHQ-1-3’(SEQ ID NO：
8)；
(5) the fluorescence probe A2V-Prob of specific detection A2V-F and A2V-R amplified productions is：
A2V-Prob:5’6-ROX-CCTCGGCCCCACTGACCCTC-3’BHQ-1(SEQ ID NO：9).
(6) the fluorescence probe GAPDH-Prob of specific detection GAPDH-F and GAPDH-R amplified productions is：
GAPDH-Prob:5’-CY5-AAAGAGCTAGGAAGGACAGGCAACTTGGC-BHQ2-3’(SEQ ID NO：
10)。
The sequence of above-mentioned reference gene GAPDH fragments referring to (Zimmermann B, Holzgreve W, Wenzel F,
Hahn S.Novel real-time quantitative PCR test for trisomy 21.Cl in
Chem.2002Feb；48(2):362-3.), it is specific as follows：
CCCCACACACATGCACTTACCTGTGCTCCCACTCCTGATTTCTGGAAAAGAGCTAGGAAGGACAGGCAACTTGGCAA
(SEQ ID NO：11).
(7) other constituents：
GoldStar Taq DNA Polymerase and buffer solution supporting GoldStar Taq DNAPolymerase
It is century to be purchased from health with dNTPs, MgCl2Purchased from Life Technology.
PCR reaction systems are prepared by table 1 below：
Table 1：(mM represents mmol/L, μM expression μm ol/L)
2nd, implementation：
PCR reaction instruments are Bio-Rad real time thermocyclers CFX96.PCR response procedures are：95 DEG C of pre-degenerations 10
Minute, then 95 DEG C 15 seconds, 60 DEG C are annealed 60 seconds, 40 circulations, in 60 DEG C of annealing steps end collection fluorescence signals.
Sample process：Extracted using Lab-Aid DNAmini extraction kid (Bio-V, Xiamen) kit
DNA, it is standby that 20~200ng/ μ L are diluted to distilled water.DNA sample can also use other conventional DNA extraction method extractions.
Pattern detection：By 2 parts of known type sample to be checked, (wherein 1 part contains HBA2 to be known:c.301-
24delinsCTCGGCCC genotype sample to be checked (abbreviation 1# samples)；Another contains-α to be known21.9Genotype treats sample
This (abbreviation 2# samples)), and 1 part of normal genotype (α α/α α, N/N) sample (abbreviation 3# samples), according to above-mentioned reaction system
And response procedures, in carrying out augmentation detection on quantitative real time PCR Instrument, record Cq values.
3rd, samples sources：All sample standard deviation sources conventional Gap-PCR technologies of hanging oneself determine the DNA sample of genotype.
4th, data analysis and result judgement：The analysis software sentence read result carried according to real-time fluorescence PCR instrument.If
CY5 passages have Cq value readings, then it represents that the sample is normally expanded；
On the premise of CY5 passages have reading, if ROX passages have reading, then it represents that the sample carries HBA2:c.301-
24delinsCTCGGCCC allele；
On the premise of CY5 passages have reading, if ROX passages do not have Cq value readings, then it represents that the sample does not carry HBA2:
C.301-24delinsCTCGGCCC allele；
On the premise of CY5 passages have reading, if FAM passages have Cq value readings, then it represents that the sample carrying-α21.9Equipotential
Gene；
On the premise of CY5 passages have reading, if FAM passages do not have Cq value readings, then it represents that the sample does not carry-α21.9
Allele.
Analyze after testing, the CY5 passages of 1# samples there are a Cq values and ROX passages there are Cq values (amplification curve is as shown in figure 1, Cq
It is worth reading as shown in Table 2), represent that 1# samples carry HBA2:C.301-24delinsCTCGGCCC allele；2# samples
CY5 passages have a Cq values and FAM passages have Cq values (amplification curve as shown in Fig. 2 Cq values reading as shown in Table 3), represent 2# samples
Carrying-α21.9Allele；The CY5 passages of 3# samples have Cq values and ROX passages and FAM passages do not have Cq values (amplification curve are such as
Shown in Fig. 3, Cq values reading is as shown in Table 4), represent that 3# samples do not carry HBA2:C.301-24delinsCTCGGCCC equipotential base
Because also not carrying-α21.9Allele.
Table 2：
Table 3：
Table 4：
It can be seen that using kit progress-α of the present invention21.9It is as a result accurate when deletion form and α 2 become heteroallele detection
Really；And wild type sample specific C q values nothing but, there is higher specificity.
Embodiment 2：Detection results of the kit of the present invention in random gene pattern sheet.
1st, the composition of kit：
With embodiment 1.
2nd, implementation：
With embodiment 1.
3rd, samples sources：
Sample (is provided) for 5 random samples by healthcare hospital for women ＆ children of Qinzhou City, and 20~200ng is diluted to distilled water, is made
For sample to be checked), and 1 through Gap-PCR and sequence verification wild type sample, 1 carrying HBA2:c.301-
24delinsCTCGGCCC allele samples, 1 carrying-α21.9Allele sample.
4th, data analysis and result judgement：
The analysis software sentence read result carried according to real-time fluorescence quantitative PCR instrument；When CY5 passages have Cq value readings
When, then the sample is normally expanded；
On the premise of CY5 passages have reading, if ROX passages have reading, then it represents that the sample carries HBA2:c.301-
24delinsCTCGGCCC allele；
On the premise of CY5 passages have reading, if ROX passages do not have Cq value readings, then it represents that the sample does not carry HBA2:
C.301-24delinsCTCGGCCC allele；
On the premise of CY5 passages have reading, if FAM passages have Cq value readings, then it represents that the sample carrying-α21.9Equipotential
Gene；
On the premise of CY5 passages have reading, if FAM passages do not have Cq value readings, then it represents that the sample does not carry-α21.9
Allele.
Table 5：
Note：Neg Ctrl=negative controls in sample type, Pos Ctrl=positive controls, Unkn=unknown sample classes
Type, in testing result, N/A=is without Cq values.
As a result show, wild type sample and positive sample detection Cq values are consistent with theory, and 1 carrying is detected in random sample
HBA2:C.301-24delinsCTCGGCCC allele.
Claims (2)
1. thalassemia-α can be detected simultaneously based on hydrolysis probes method21.9The kit that deletion form and α 2 make a variation, including amplification
Primer and fluorescence probe, it is characterised in that：
The variations of α 2 are the mutant gene type of the genes of α 2 in α-globin gene cluster NG_000006.1, and the genotype is described
For HBA2:c.301-24delinsCTCGGCCC；
Described amplimer is：- α in α-globin gene cluster can be expanded a pair21.9The primer of the characteristic sequence of allele
21.9-F and 21.9-R, can expand HBA2 in α-globin gene cluster a pair:C.301-24delinsCTCGGCCC equipotential base
The primer A2V-F and A2V-R of the characteristic sequence of cause, and the primer GAPDH-F of reference gene GAPDH fragments can be expanded for a pair
And GAPDH-R；
Described fluorescence probe is：The fluorescence probe 21.9-Prob of one specific detection 21.9-F and 21.9-R amplified production,
The fluorescence probe A2V-Prob, a specific detection GAPDH-F of one specific detection A2V-F and A2V-R amplified production and
The fluorescence probe GAPDH-Prob of GAPDH-R amplified productions；
Wherein：
It is described to expand-α in α-globin gene cluster21.9In the primer pair of the characteristic sequence of allele,
21.9-F：5’-GGAGCTTTTCCTTCCCTGGAACG-3’；
21.9-R：5’-TGTGGTTGGAGAATGGAGGTGG-3’；
It is described to expand HBA2 in α-globin gene cluster：C.301-24delinsCTCGGCCC the characteristic sequence of allele
Primer pair in,
A2V-F：5’-GGGTTGCGGGAGGTGTAG-3’；
A2V-R：5’-TCCTTGGTCTGAGACAGGTA-3’；
In the primer pair that reference gene GAPDH fragments can be expanded,
GAPDH-F：5’-CCCCACACACATGCACTTACC-3’；
GAPDH-R：5’-CCTAGTCCCAGGGCTTTGATT-3’；
The fluorescence probe 21.9-Prob of the specific detection 21.9-F and 21.9-R amplified productions is：
21.9-Prob：5’-6-FAM-CTGGGGAAGGGTGGGTGGGAATAACAGC-BHQ-1-3’；
The fluorescence probe A2V-Prob of described specific detection A2V-F and A2V-R amplified production is：
A2V-Prob：5’-6-ROX-CCTCGGCCCCACTGACCCTC-BHQ-1-3’；
The fluorescence probe GAPDH-Prob of described specific detection GAPDH-F and GAPDH-R amplified production is：
GAPDH-Prob：5’-CY5-AAAGAGCTAGGAAGGACAGGCAACTTGGC-BHQ-2-3’.
2. kit according to claim 1, it is characterised in that：Described kit also includes buffer solution, enzyme liquid, MgCl2
And dNTP.
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| CN107267648B (en) * | 2017-08-04 | 2020-04-14 | 亚能生物技术（深圳）有限公司 | Kit for rapidly detecting common mutant genes of thalassemia |
| CN107245528B (en) * | 2017-08-04 | 2020-04-21 | 亚能生物技术（深圳）有限公司 | Rapid detection kit for common thalassemia mutant genes of Chinese population |
| CN114150051B (en) * | 2021-11-05 | 2024-04-02 | 上海源赏生物科技有限公司 | Kit and method for comprehensively detecting five complex genetic diseases in integrated manner |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103255225A (en) * | 2013-05-27 | 2013-08-21 | 钦州市妇幼保健院 | Thalassemia gene detection method based on fluorescence labeling quantitation PCR (Polymerase Chain Reaction) technology |
| EP2633067A1 (en) * | 2010-10-27 | 2013-09-04 | CapitalBio Corporation | Luminophore-labeled molecules coupled with particles for microarray-based assays |
| CN103966319A (en) * | 2014-04-16 | 2014-08-06 | 龙驹 | Kit for detecting common deletion form alpha-thalassemia and using method of kit |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP2633067A1 (en) * | 2010-10-27 | 2013-09-04 | CapitalBio Corporation | Luminophore-labeled molecules coupled with particles for microarray-based assays |
| CN103255225A (en) * | 2013-05-27 | 2013-08-21 | 钦州市妇幼保健院 | Thalassemia gene detection method based on fluorescence labeling quantitation PCR (Polymerase Chain Reaction) technology |
| CN103966319A (en) * | 2014-04-16 | 2014-08-06 | 龙驹 | Kit for detecting common deletion form alpha-thalassemia and using method of kit |
Non-Patent Citations (2)
| Title |
|---|
| 《Detection of three common α-thalassemia in non-deletion types and six common thalassemia in deletion types by QF-PCR》;Ju Long等;《Clinical Biochemistry》;20131231;第46卷(第18期);1860-1864页 * |
| 《多重荧光定量PCR快速检测缺失型α地中海贫血》;彭建明等;《广东医学》;20130430;第34卷(第8期);1195-1196页 * |
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