CN202830036U - Real-time fluorescence quantification polymerase chain reaction (PCR) reagent kit for detecting Y-chromosome microdeletion - Google Patents
Real-time fluorescence quantification polymerase chain reaction (PCR) reagent kit for detecting Y-chromosome microdeletion Download PDFInfo
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- CN202830036U CN202830036U CN201220447631.4U CN201220447631U CN202830036U CN 202830036 U CN202830036 U CN 202830036U CN 201220447631 U CN201220447631 U CN 201220447631U CN 202830036 U CN202830036 U CN 202830036U
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Abstract
The utility model discloses a real-time fluorescence quantification polymerase chain reaction (PCR) reagent kit for detecting Y-chromosome microdeletion. The reagent kit comprises a kit body, PCR reaction liquid and at least one primer probe combination arranged on the kit body. The primer probe combination comprises one or more of primer probe combinations which are an SY 84 primer probe combination used for detecting an SY 84 locus, an SY 86 primer probe combination used for detecting an SY 86 locus, an SY 127 primer probe combination used for detecting an SY 127 locus, an SY 134 primer probe combination used for detecting an SY 134 locus, an SY 254 primer probe combination used for detecting an SY 254 locus and an SY 255 primer probe combination used for detecting an SY 255 locus. According to the reagent kit, whether deletion of a plurality of locuses of an AZF zone of a Y-chromosome happens can be specifically and effectively detected in reaction liquid, and accordingly time and cost can be effectively reduced.
Description
Technical field
The utility model relates to a kind of micro-deleted test kit of Y chromosome that detects.
Background technology
According to statistics, the whole world has 15% Mr. and Mrs to suffer from sterile (pregnant) disease, wherein more than 50% by due to the bridegroom's or husband's side.Recent study shows that the male sterility more than 30% is because the dyszoospermia that inherited genetic factors causes, be usually expressed as male Y chromosome long-armed (q arm) 11 districts upper without the smart factor gene (Azoospermiafactor of family, AZF) deletion mutantion in a plurality of sites, wherein the micro-deleted of any one seat all can cause dyszoospermia, and can't treat for the male sterility that causes because Y chromosome is micro-deleted.Y chromosome is micro-deleted can for clinical treatment provides guidance, reduce unnecessary economic waste in detection before intracytoplasmic sperm injection (ICSI) technology assisted reproduction treatment.The micro-deleted examination project of classifying male sterility as of Y chromosome has been advised detecting by the relevant association of Europe, the U.S. and China at present.
Detect at present a lot, the most frequently used PCR-RFLP method of method of gene microdeletion, also have in addition reversal point hybrid method, allele-specific polymerase chain reaction, direct sequencing etc.The latter is methodical gold standard, but owing to its somewhat expensive, is not widely used.Although and PCR-RFLP method expense is lower, simple to operate, specificity and susceptibility are not ideal, and it is little that it detects flux, inapplicable for the clinical detection that sample size is larger.
The utility model content
The utility model provide a kind of can effectively reduce time and cost for detection of the micro-deleted test kit of Y chromosome.
It is a kind of for detection of the micro-deleted real-time fluorescent PCR reagent case of Y chromosome that the utility model provides, comprise box body, liner, the first container of PCR reaction solution is housed and at least one second container of primer probe combinations is housed, described liner has a plurality of container holes, described the first container places the described container hole corresponding with it, described second container places the described container hole corresponding with it, and described second container comprises the second container that the SY84 primer probe combinations that detects the SY84 site is housed, the second container of the SY86 primer probe combinations that detects the SY86 site is housed, the second container of the SY127 primer probe combinations that detects the SY127 site is housed, the second container of the SY134 primer probe combinations that detects the SY134 site is housed, the second container of the SY254 primer probe combinations that detects the SY254 site is housed and one or more in the second container of the SY255 primer probe combinations that detects the SY255 site are housed.
The micro-deleted real-time fluorescent PCR reagent case of described detection Y chromosome comprises also being equipped with and detects the 3rd container that male gender determines the SRY primer probe combinations of gene SRY that described the 3rd container places the described container hole corresponding with it.
Described second container comprises the second container that the SY84 primer probe combinations that detects the SY84 site is housed, the second container of the SY86 primer probe combinations that detects the SY86 site is housed, the second container of the SY127 primer probe combinations that detects the SY127 site is housed, the second container of the SY134 primer probe combinations that detects the SY134 site is housed, the second container of the SY254 primer probe combinations that detects the SY254 site is housed and a plurality of in the second container of the SY255 primer probe combinations that detects the SY255 site are housed.。
Described second container comprises the second container that the SY84 primer probe combinations that detects the SY84 site is housed, the second container of the SY86 primer probe combinations that detects the SY86 site is housed, the second container of the SY127 primer probe combinations that detects the SY127 site is housed, the second container of the SY134 primer probe combinations that detects the SY134 site is housed, the second container of the SY254 primer probe combinations that detects the SY254 site is housed and the second container of the SY255 primer probe combinations that detects the SY255 site is housed.
The micro-deleted real-time fluorescent PCR reagent case of described detection Y chromosome also comprises the 4th container that positive reference substance is housed and the 5th container that the negative control product are housed, and described the 4th container and the 5th container place respectively the described container hole corresponding with it.
Described SY84, SY86, SY127, SY134, SY254 and SY255 primer probe combinations include upstream primer, downstream primer and Taqman fluorescent probe.
Different primer probe combinations can select to detect one or more site as required for detection of the different loci in Y chromosome AZF zone, corresponding, select the primer probe combinations corresponding with this site.
The AZF zone of Y chromosome forms by AZFa is regional, AZFb is regional and AZFc is regional.
Described test kit also comprises the SRY primer probe combinations that determines gene SRY for detection of male gender, and described SRY primer probe combinations comprises upstream primer, downstream primer and Taqman fluorescent probe.
SRY primer probe combinations is used for Quality Control, and it can select to adopt or do not adopt according to testing requirement.
Described primer probe combinations is comprised of SY84, SY86, SY127, SY134, SY254 and SY255 primer probe combinations, the Tm value differs and fluctuates in 2-4 ℃ between the described primer, the Tm value differs and fluctuates in 2-4 ℃ between the described probe, and to differ be about 10 ℃ to the Tm value between the primer probe.
Primer comprises for the primer in each site with for the primer of gene SRY.Probe comprises for the probe in each site with for the probe of gene SRY.
Described PCR reaction solution also comprises the 10 times of damping fluids (10*Buffer) that contain MgCl, KCl, Tris, the dNTP that contains dATP, dCTP, dGTP, dUTP and rTaq enzyme.
Wherein, MgCl concentration is 1.5mM, and KCl concentration is that 60mM, Tris concentration are 12mM.
Wherein, dNTP concentration is 0.2mM.
Wherein, synthetic primer concentration is that 300uM, concentration and probe concentration are 100uM.
A kind of for detection of the micro-deleted method of Y chromosome, comprise the steps:
A) from people's anticoagulation, extract genomic dna;
B) take described genomic dna sample as template, utilize SY84 claimed in claim 7, SY86, SY127, SY134, SY254 and SY255 primer probe combinations to carry out pcr amplification and detection;
C) according to the color of the corresponding probe in detecting passage of fluorescence probe Marker selection;
D) have or not the AZF zone of judging corresponding Y chromosome whether to have disappearance according to amplification curve color and curve.
Wherein, the method adopts the quadruple quantitative fluorescent PCR to divide two pipes that 6 sites, 3 zones of Y chromosome are detected.
The beneficial effects of the utility model are: utilize this test kit, whether a plurality of sites that are can be in reaction solution special and that detect efficiently the AZF zone of Y chromosome lack, thereby can effectively reduce time and cost.
Description of drawings
Fig. 1 is the structural representation of present embodiment test kit;
Fig. 2-the 11st shows the amplified fluorescence graphic representation of present embodiment detected result, wherein,
4 curves of Fig. 2 represent respectively the amplification curve in SRY, SY84, SY127 and SY254 site;
4 curves of Fig. 3 represent respectively the amplification curve in SRY, SY134, SY255 and SY86 site;
3 curves of Fig. 4 represent respectively the amplification curve in SRY, SY127 and SY254 site;
3 curves of Fig. 5 represent respectively the amplification curve in SRY, SY134 and SY255 site;
3 curves of Fig. 6 represent respectively the amplification curve in SRY, SY84 and SY254 site;
4 curves of Fig. 7 represent respectively the amplification curve in SRY, SY86, SY134 and SY255 site;
3 curves of Fig. 8 represent respectively the amplification curve in SRY, SY86 and SY255 site;
4 curves of Fig. 9 represent respectively the amplification curve in SRY, SY84, SY127 and SY254 site;
3 curves of Figure 10 represent respectively the amplification curve in SRY, SY84 and SY127 site;
3 curves of Figure 11 represent respectively the amplification curve in SRY, SY86 and SY134 site.
Embodiment
The utility model use the quadruple quantitative fluorescent PCR divide two pipes to Y chromosome AZFa zone the SY84 site and the SY86 site, to SY127 site and the SY134 site in AZFb zone, AZFc SY254 site, zone and SY255 site are detected, detect simultaneously male gender and determine that gene SRY is as Quality Control, normally to have educated male sex's dna sample as positive control, women's dna sample is as negative control, and aqua sterilisa prevents that as blank the check system is contaminated.
As shown in Figure 1, present embodiment detects the micro-deleted real-time fluorescent PCR reagent case of Y chromosome, comprise box body 1, liner 2, the first container 4 of PCR reaction solution is housed and at least one second container 3 of primer probe combinations is housed, described liner 2 has a plurality of container holes, described the first container 4 places the described container hole corresponding with it, described second container 3 places the described container hole corresponding with it, and described second container 3 comprises the second container that the SY84 primer probe combinations that detects the SY84 site is housed, the second container of the SY86 primer probe combinations that detects the SY86 site is housed, the second container of the SY127 primer probe combinations that detects the SY127 site is housed, the second container of the SY134 primer probe combinations that detects the SY134 site is housed, the second container of the SY254 primer probe combinations that detects the SY254 site is housed and one or more in the second container of the SY255 primer probe combinations that detects the SY255 site are housed.
This test kit can also comprise the 3rd container 5 that the SRY primer probe combinations that detects male gender decision gene SRY is housed, and described the 3rd container 5 places the described container hole corresponding with it.
Primer probe combinations for detection of SY84 site, Y chromosome AZFa zone is defined as SY84 primer probe combinations, it comprises upstream primer, downstream primer and Taqman fluorescent probe, fluorescent probe is with one of FAM, HEX, CY5, ROX mark 5 ' end, its fluorescent mark is identical with the SY86 site, but different with the four pairs of probes in AZFb, AZFc zone and SRY probe.But the probe in above-mentioned SY84 site, upstream and downstream primer random combine.
Primer probe combinations for detection of SY86 site, Y chromosome AZFa zone is defined as SY86 primer probe combinations, it comprises upstream primer, downstream primer and Taqman fluorescent probe, fluorescent probe is with one of FAM, HEX, CY5, ROX mark 5 ' end, its fluorescent mark is identical with the SY84 site, but different with the four pairs of probes in AZFb, AZFc zone and SRY probe.But the probe in above-mentioned SY86 site, upstream and downstream primer random combine.
Primer probe combinations for detection of SY127 site, Y chromosome AZFb zone is defined as SY127 primer probe combinations, it comprises upstream primer, downstream primer and Taqman fluorescent probe, fluorescent probe is with one of FAM, HEX, CY5, ROX mark 5 ' end, its fluorescent mark is identical with the SY134 site, but different with the four pairs of probes in AZFa, AZFc zone and SRY probe.But the probe in above-mentioned SY127 site, upstream and downstream primer random combine.
Primer probe combinations for detection of SY134 site, Y chromosome AZFb zone is defined as SY134 primer probe combinations, it comprises upstream primer, downstream primer and Taqman probe, fluorescent probe is with one of FAM, HEX, CY5, ROX mark 5 ' end, its fluorescent mark is identical with the SY127 site, but different with the four pairs of probes in AZFa, AZFc zone and SRY probe.But the probe in above-mentioned SY134 site, upstream and downstream primer random combine.
Primer probe combinations for detection of SY254 site, Y chromosome AZFc zone is defined as SY254 primer probe combinations, it comprises upstream primer, downstream primer and Taqman fluorescent probe, fluorescent probe is with one of FAM, HEX, CY5, ROX mark 5 ' end, its fluorescent mark is identical with the SY255 site, but different with the four pairs of probes in AZFa, AZFb zone and SRY probe.But the probe in above-mentioned SY254 site, upstream and downstream primer random combine.
Primer probe combinations for detection of SY255 site, Y chromosome AZFc zone is defined as SY255 primer probe combinations, it comprises upstream primer, downstream primer and Taqman fluorescent probe, fluorescent probe is with one of FAM, HEX, CY5, ROX mark 5 ' end, its fluorescent mark is identical with the SY254 site, but different with the four pairs of probes in AZFa, AZFb zone and SRY probe.But the probe in above-mentioned SY255 site, upstream and downstream primer random combine.
The primer probe combinations that determines gene SRY for detection of male gender is defined as SRY primer probe combinations, it comprises upstream primer, downstream primer and Taqman fluorescent probe, fluorescent probe is with one of FAM, HEX, CY5, ROX mark 5 ' end, and its fluorescent mark and AZFa, the regional 6 pairs of probes of AZFb, AZFc are different.But the probe of said gene SRY, upstream and downstream primer random combine.
Whether the AZFa zone that above-mentioned SY84, SY86 primer probe combinations are used for the specific detection Y chromosome exists disappearance.
Whether the AZFb zone that above-mentioned SY127, SY134 primer probe combinations are used for the specific detection Y chromosome exists disappearance.
Whether the AZFc zone that above-mentioned SY254, SY255 primer probe combinations are used for the specific detection Y chromosome exists disappearance.
Above-mentioned SRY primer is visited this combination and is used for Quality Control, whether normally detects the PCR process.
Above-mentioned all sequences can be replaced by its complementary base sequences thereof.
Present embodiment also comprises the required reagent of pcr amplification for detection of the micro-deleted test kit of male Y chromosome: 10*Buffer(contains the buffered soln of magnesium ion), dNTP(contains dATP, dCTP, dGTP, dUTP), the rTaq enzyme.
For detection of the micro-deleted test kit of Y chromosome, MgCl concentration is 1.5mM among the 10*Buffer, and KCl concentration is that 60mM, Tris concentration are 12mM.
For detection of the micro-deleted test kit of Y chromosome, dNTP concentration is 0.2mM.
All synthetic primer concentration are 300uM, and concentration and probe concentration is 100uM.
Tm(melting temperature between the described primer, the melting temperature(Tm) of oligonucleotide) value differs and fluctuates in 2-4 ℃, and the Tm value differs and fluctuates in 2-4 ℃ between the probe, and to differ be about 10 ℃ to the Tm value between the primer probe.
A kind of for detection of the micro-deleted gene tester of Y chromosome, may further comprise the steps:
A) from people's anticoagulation, extract genomic dna;
B) take step a income earner genomic dna sample as template, utilize each above-mentioned primer probe combinations that the AZF zone of Y chromosome and SRY are carried out specific PCR and detect;
C) set the PCR reaction conditions, according to the color of the corresponding probe in detecting passage of fluorescence probe Marker selection;
D) have or not the no existence disappearance in 6 sites, judgement corresponding A ZF zone according to amplification curve color and curve.
The PCR reaction conditions is: 94 ℃ of 2min; 94 ℃ of 5S, 52 ℃ of 45S, 60 ℃ of 45S, this process is carried out 40 circulations.
It is micro-deleted to specific detection Y chromosome AZFa, AZFb, AZFc zone that present embodiment has designed 6 groups of probes, 1 group of male gender determines that gene probe is to carrying out Quality Control, 6 groups of probes are to adhering to separately in two PCR reactions in addition, carry out PCR under the identical conditions, the negative contrast of women's dna sample, aqua sterilisa are that blank is contaminated to prevent the check system.
One, material
1. instrument: real time fluorescent quantitative instrument
Probe, design of primers: with above-mentioned 6 pairs of probes, the specific detection of the primer pair AZF sequence of having designed, and design simultaneously 1 pair of probe, primer as Quality Control, detect male gender and determine gene, whether normally carry out to detect the PCR testing process, normally to have educated male sex's dna sample as positive control, women's dna sample is as negative control, and aqua sterilisa prevents that as blank the check system is contaminated.
2. use following probe primer to detecting AZFa, AZFb, site, AZFc zone disappearance.
SY84 upstream primer 5'-CCTATTTGTTTTAAGGTGCCATTCC-3'
SY84 downstream primer 5'-GCTTGCCTGAGCATCCTACAG-3'
SY84 probe 5'-VIC-CTCTACCTCCTTCCCCCAGTGCCCA-BHQ-3'
SY86 upstream primer 5'-GGCTTCCCAGAGTTGTTACAAAG-3'
SY86 downstream primer 5'-CTCAAGGACTGTGAGAATCAAGGTT-3'
SY86 probe 5'-VIC-AATCCCAAAGACTGGGCCCCTTAAACA-BHQ-3'
SY127 upstream primer 5'-GGCTCACAAACGAAAAGAAAAAG-3'
SY127 downstream primer 5'-CTTTTTGTGGGCATGAACACA-3'
SY127 probe 5'-FAM-TAGCACCCACTGGAATCTACCAAAGCCC-BHQ-3'
SY134 upstream primer 5'-GACTCACTATAGGGCGAATTGG-3'
SY134 downstream primer 5'-GGTGAGGCAGACAGTTTTGTAG-3'
SY134 probe 5'-FAM-TACGGGCCCCCCCTCGAG-BHQ-3'
SY254 upstream primer 5'-AAGGCAAAATCGTGCCAAAC-3'
SY254 downstream primer 5'-CCATTGTTCATGATGTATGTTAAGGTAA-3'
The SY254 probe
5'-CTGTTTTTGTTGGTGGAATTGATGCTAGGG-BHQ-3'
SY255 upstream primer 5'-GTTACAGGATTCGGCGTGAT-3'
SY255 downstream primer 5'-CTCGTCATGTGCAGCCAC-3'
SY255 probe 5'-ROX-TAGGTTTCAGTGTTTGGATTCCGCCA-BHQ-3'
SRY upstream primer 5'-TTTTCGGCTTCAGTAAGCATTTT-3'
The SRY downstream primer
5'-AATCCCAGAATGCGAAACTCA-3'
SRY probe 5'CY5-CACTGGTATCCCAGCTGCTTGCTGATC-BHQ-3'
3.PCR reaction reagent: 10*Buffer(contains magnesium ion), dNTP(contains dATP, dCTP, dGTP, dUTP), the rTaq enzyme.
Two, method
1. extracting genome DNA
Buy test kit or routinely molecular biology method extract genomic dna.Extract respectively 1 routine normal male genomic dna, 4 routine improper male gene group DNA and 1 routine women's genomic dna.
2. fluorescence quantitative PCR detection
The PCR reaction is front to be put on ice naturally dissolving with reagent, get the PCR pipe and carry out mark, add respectively aqua sterilisa, 10*Buffer, dNTP, rTaq enzyme, dna sample to be detected, each sample carries out simultaneously 2 PCR and detects, be PCR reaction solution A 22.5 μ l+ sample DNA 2.5 μ l to be detected, the same sample DNA 2.5 μ l to be detected of PCR reaction solution B22.5 μ l+, the PCR reaction system sees Table 2.Positive control (normal male genomic dna) and negative control (women's genomic dna), aqua sterilisa blank are all set up in each detection.
Table 1PCR reaction system
Detect by following PCR reaction conditions: 94 ℃ of 2min; 94 ℃ of 5S, 52 ℃ of 45S, 60 ℃ of 45S, this process is carried out 40 circulations.
Three, interpretation of result:
1. detection success conditions: after quantitative fluorescent PCR finishes, four looks level and smooth amplification curve all appears in the positive control a, the b that add reaction solution A and B, knee point is clear, the whole collimation good (shown in Fig. 1-10) of amplification curve, and level and smooth amplification curve does not appear in negative control c.Curve C T value fluctuates normal for detecting between 18-30.
2. select sense channel and color according to institute's different probe fluorescent mark, judge that according to different passage amplification situations corresponding site has or not disappearance.
3. amplification all should appear in all sample SRY probes to be detected, otherwise is the DNA extraction failure.
Fig. 2-3 is the normal male detected result, amplification appears in this sample SRY probe, show this pattern detection success, amplification does not appear in negative control, blank, amplification all appears in 7 probes of positive control, show that testing process is error free, pollution-free, amplification all appears in all detection site of this sample, and is micro-deleted without Y chromosome.
Fig. 4-5 is same sterile male sex's sample, amplification appears in this sample SRY probe, show this pattern detection success, amplification does not appear in negative control, blank, amplification all appears in 7 probes of positive control, show that testing process is error free, pollution-free, SY127, SY134, SY254, SY255 site amplification occurs and show that this sample AZFb, AZFc zone is without disappearance, amplification does not appear in SY84, SY86 site, there is disappearance in the SY84, the SY86 site that show the AZFa zone, wherein Fig. 4 shows SY84 site disappearance, and Fig. 5 shows SY86 site disappearance.
Fig. 6-7 is same sterile male sex's sample, and amplification appears in this sample SRY probe, shows this pattern detection success, and amplification does not appear in negative control, blank, and amplification all appears in 7 probes of positive control, shows that testing process is error free, pollution-free.Amplification all appears in SY84, SY86, SY134, SY254, SY255 site, and amplification does not appear in the SY127 site, shows the SY127 site disappearance in AZFb zone.Wherein, Fig. 7 shows SY127 site disappearance.
Fig. 8-9 is same sterile male sex's sample, and amplification appears in this sample SRY, shows this pattern detection success, and amplification appears in SY84, SY86, SY127, SY254, SY255, and amplification does not appear in the SY134 site, shows the SY134 site disappearance in AZFb zone.
Figure 10-11 is same sterile male sex's sample, and amplification appears in this sample SRY, shows this pattern detection success, and amplification appears in SY84, SY86, SY127, SY134, and amplification does not appear in SY254, SY255, shows SY254, the SY255 site disappearance in AZFc zone.Wherein, Figure 10 shows SY254 site disappearance, and Figure 11 shows SY255 site disappearance.
The purpose of this utility model is to provide utilizes real-time fluorescence quantitative PCR in conjunction with the Taqman probe the micro-deleted site of Y chromosome to be detected.Thereby real-time fluorescence quantitative PCR is realized starting template quantitatively and is qualitatively analyzed by the real-time collecting of pcr amplification to each circulation products fluorescent signal.This technology has reduced the error in the regular-PCR troublesome operation process, has reduced the possibility of PCR product pollution, have simple to operate, quick, detect that flux is high, susceptibility, characteristics that specificity is high, be a kind of Protocols in Molecular Biology that should be widely promoted.The utility model uses the quadruple quantitative fluorescent PCR to divide two pipes to SY84 site, Y chromosome AZFa zone, SY86 site, SY127 site, AZFb zone, SY134 site, SY254 site, AZFc zone, SY255 site are detected, detect simultaneously male gender and determine that gene SRY is as Quality Control, normally to have educated male sex's dna sample as positive control, women's dna sample is as negative control, and aqua sterilisa prevents that as blank the check system is contaminated.
Above content is in conjunction with concrete embodiment further detailed description of the utility model, can not assert that implementation of the present utility model is confined to these explanations.For the utility model person of an ordinary skill in the technical field, without departing from the concept of the premise utility, can also make some simple deduction or replace.
Claims (5)
1. one kind is detected the micro-deleted real-time fluorescent PCR reagent case of Y chromosome, it is characterized in that, comprise box body, liner, the first container of PCR reaction solution is housed and at least one second container of primer probe combinations is housed, described liner has a plurality of container holes, described the first container places the described container hole corresponding with it, described second container places the described container hole corresponding with it, and described second container comprises the second container that the SY84 primer probe combinations that detects the SY84 site is housed, the second container of the SY86 primer probe combinations that detects the SY86 site is housed, the second container of the SY127 primer probe combinations that detects the SY127 site is housed, the second container of the SY134 primer probe combinations that detects the SY134 site is housed, the second container of the SY254 primer probe combinations that detects the SY254 site is housed and one or more in the second container of the SY255 primer probe combinations that detects the SY255 site are housed.
2. the micro-deleted real-time fluorescent PCR reagent case of detection Y chromosome as claimed in claim 1, it is characterized in that, also comprise being equipped with and detect the 3rd container that male gender determines the SRY primer probe combinations of gene SRY, described the 3rd container places the described container hole corresponding with it.
3. the micro-deleted real-time fluorescent PCR reagent case of detection Y chromosome as claimed in claim 2, it is characterized in that, described second container comprises the second container that the SY84 primer probe combinations that detects the SY84 site is housed, the second container of the SY86 primer probe combinations that detects the SY86 site is housed, the second container of the SY127 primer probe combinations that detects the SY127 site is housed, the second container of the SY134 primer probe combinations that detects the SY134 site is housed, the second container of the SY254 primer probe combinations that detects the SY254 site is housed and a plurality of in the second container of the SY255 primer probe combinations that detects the SY255 site are housed.。
4. the micro-deleted real-time fluorescent PCR reagent case of detection Y chromosome as claimed in claim 3, it is characterized in that, described second container comprises the second container that the SY84 primer probe combinations that detects the SY84 site is housed, the second container of the SY86 primer probe combinations that detects the SY86 site is housed, the second container of the SY127 primer probe combinations that detects the SY127 site is housed, the second container of the SY134 primer probe combinations that detects the SY134 site is housed, the second container of the SY254 primer probe combinations that detects the SY254 site is housed and the second container of the SY255 primer probe combinations that detects the SY255 site is housed.
5. such as the micro-deleted real-time fluorescent PCR reagent case of the described detection Y chromosome of any one among the claim 1-4, it is characterized in that, also comprise the 4th container that positive reference substance is housed and the 5th container that the negative control product are housed, described the 4th container and the 5th container place respectively the described container hole corresponding with it.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014036925A1 (en) * | 2012-09-04 | 2014-03-13 | Luo Ziyi | Real-time fluorescence quantitative pcr kit and method for detecting y-chromosome microdeletion |
| CN105132530A (en) * | 2015-07-22 | 2015-12-09 | 广州市达晖生物技术有限公司 | Kit for rapid detection of human Y chromosomal microdeletion |
-
2012
- 2012-09-04 CN CN201220447631.4U patent/CN202830036U/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014036925A1 (en) * | 2012-09-04 | 2014-03-13 | Luo Ziyi | Real-time fluorescence quantitative pcr kit and method for detecting y-chromosome microdeletion |
| CN105132530A (en) * | 2015-07-22 | 2015-12-09 | 广州市达晖生物技术有限公司 | Kit for rapid detection of human Y chromosomal microdeletion |
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