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Method, primer, probe, and fluorescence PCR (polymerase chain reaction) kit for detecting gender of fetus in vitro

An in vitro detection, fetal technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc.

Inactive Publication Date: 2014-03-19
叶盛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the extremely low content of ffDNA in maternal blood and the very high content of maternal background DNA, the normal PCR amplification method, due to the influence of maternal background DNA and the incomplete stringency of DNA double-strand matching, led to poor test results. low accuracy

Method used

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  • Method, primer, probe, and fluorescence PCR (polymerase chain reaction) kit for detecting gender of fetus in vitro
  • Method, primer, probe, and fluorescence PCR (polymerase chain reaction) kit for detecting gender of fetus in vitro

Examples

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Embodiment 1

[0029] 1. Sample processing: Samples 1-3 are blood drawn from the veins of pregnant women at 6 weeks of gestation, and samples 4-6 are blood drawn from the veins of pregnant women at 12 weeks of pregnancy. Samples 1-6 were centrifuged to remove blood cells, plasma was collected, and plasma DNA was extracted.

[0030] 2. Design and synthesis of primers and probes: The DNA sequence of SPY gene was retrieved from GeneBank, and then compared by DNAStar software, and primer sequences and corresponding PNA sequences were designed and synthesized in relatively conserved regions. The fluorescent group FAM (emission wavelength: 522nm) is labeled on the glutamic acid end of the PNA probe, and the quenching group DABSYL is labeled on the lysine end.

[0031] 3. PCR reaction system:

[0032] Template 2 μl

[0033] PCR-MIX 2.5 μl

[0034] Taq enzyme 1μl

[0035] Upstream primer 2 μl

[0036] Downstream primer 2 μl

[0037] Probe 1μl

[0038] wxya 2 O 14.5 μl

[0039] 25 μl tot...

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Abstract

The invention discloses a method for detecting gender of fetus in vitro, which comprises the following steps: (1) centrifuging a maternal blood sample, extracting plasma and extracting the plasma DNA; (2) performing fluorescent quantitive PCR by using the extracted DNA as the template, adding a primer SRY-P1 and a probe SPY-PNA1; and (3) detecting a PCR product. The invention further discloses a primer and probe for detecting gender of fetus in vitro, and a fluorescence PCR kit for detecting the gender of the fetus in vitro. The invention provides a method for detecting the gender of the fetus in vitro at early pregnancy, and the primer SRY-P1 and the probe SPY-PNA1 are designed according to genetic information gene SRY only in men so as to perform the fluorescent quantitive PCR on the maternal blood sample. The method is used for detecting the gender of fetus in vitro, and the detection accuracy rate on the maternal pregnancy for 6 weeks can achieve 100%. The kit designed according to the method is simple to use, and the result reliability is high.

Description

technical field [0001] The invention relates to a method for detecting fetal sex in vitro by maternal blood samples, primers and probes, and a fluorescent PCR kit. Background technique [0002] Some genetic disorders associated with sex may occur only in males, while others may only occur in females. Therefore, for families who may have a history of such genetic diseases, early identification of fetal sex will be beneficial to prenatal and postnatal care programs. In the past, fetal sex determination was often performed using invasive first-trimester diagnostic methods, such as chorionic villus biopsy. Due to the direct sampling of pregnancy tissue, the accuracy rate of interventional early pregnancy diagnosis is very high. However, because the diagnostic process needs to use a needle to puncture the uterus for sampling, this operation is likely to cause damage to the mother and fetus, and in severe cases can lead to miscarriage. [0003] Ultrasound diagnosis can also be ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686
Inventor 叶盛
Owner 叶盛
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