Plasmodium nucleate type detection kit and application method thereof

A technology for detecting kits and protozoan nucleic acids, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, and resistance to vector-borne diseases, etc. It can solve unreliable diagnosis, inability to detect differences in Plasmodium species, and false negatives And other issues

Inactive Publication Date: 2019-03-19
合肥欧创基因生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, this condition can lead to false negative results or unreliable diagnoses
[0008] In addition, there is also a nested PCR detection for Plasmodium 18SrDNA. Because the Plasmodium genome is rich in AT, the primer design required for this detection method is relatively complicated, the specificity is low, and this method requires two rounds of amplification reactions. The operation is cumbersome
[0009] In short, the above-mentioned commonly used malaria diagnostic methods have problems such as limited detection, cumbersome operations, the need for well-trained experts, or the inability to detect differences in Plasmodium species; the market needs a Plasmodium nucleic acid with strong specificity and high sensitivity Typing detection kit, the present invention solves such problems

Method used

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  • Plasmodium nucleate type detection kit and application method thereof
  • Plasmodium nucleate type detection kit and application method thereof
  • Plasmodium nucleate type detection kit and application method thereof

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Embodiment Construction

[0098] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.

[0099] A Plasmodium nucleic acid typing detection kit, comprising: PCR reaction solution I containing mixed primers, PCR reaction solution II containing typing probes, Taq / UNG enzyme mixture, containing P.v, P.f, P.o, P.m mitochondrial cox3 Positive control and negative control of the recombinant plasmid of the specific region sequence of the gene;

[0100] PCR reaction solution I includes:

[0101] water;

[0102] 10×PCR buffer;

[0103] 1-25mM Mg ion;

[0104] 1-25mM dNTPs;

[0105] 1-100 μM P.v cox3 gene upstream primer VF: SEQ.ID.NO:1, CCTTACGTACTCTAGCTTTTA;

[0106] 1-100μM P.v cox3 gene downstream primer VR: SEQ.ID.NO:2, GAGTTCCTTTTTCTATTCAGAATAA;

[0107] 1-50 μM P.v cox3 gene probe VP: SEQ.ID.NO:9,

[0108] CAATATTATTTGTCTATACTAGATACTATAGTT;

[0109] 1-100 μM P.f cox3 gene upstream primer FF: SEQ.ID.NO:3, CCTTACGTACTCTAGCT...

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Abstract

The invention discloses a plasmodium nucleate type detection kit and an application method thereof. The kit comprises a PCR reaction solution I comprising a mixed primer, a PCR reaction solution II comprising a typing probe, a Taq / UNG enzyme mixed solution, and a positive reference and negative reference comprising recombinant plasmids of P.v, P.f, P.o and P.m mitochondria cox gene specific area sequence; the mixed primer comprises a primer sequence used for amplifying a P.v, P.f, P.o and P.m mitochondria cox gene specific areas, and a primer sequence for amplifying an HBB gene; and the typingprobe comprises a sequence for typing identifying the P.v, P.f, P.o and P.m mitochondria cox 3 gene pecific areas and a probe sequence for identifying the HBB gene. By adopting the design of the invention, the plasmonium in a sample can be detected, and four plasmonium can be accurately typed; and the plasmonium mitochondria cox3 gene is adopted as a target gene to perform the detection, so thatthe specificity and the sensitivity are higher.

Description

technical field [0001] The invention relates to the field of disease detection assistance, in particular to a malaria parasite nucleic acid typing detection kit and a use method thereof. Background technique [0002] Malaria, a widespread infectious disease in tropical and subtropical regions, is caused by infection by the parasitic protozoa Plasmodium. Four types of Plasmodium that can infect humans, including Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale and Plasmodium vivax. [0003] Furthermore, malaria has a negative impact on national and individual wealth. Malaria costs Africa tens of billions of dollars each year and slows its gross domestic product every year. Malaria can lead to lost productivity, driving societies deeper into poverty and negatively affecting investment and tourism. In addition, due to malaria's high mortality rate for children under the age of five, it also has a greater impact on the social structure. [0004] Diagnosing malari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156Y02A50/30
Inventor 黄希文杨芳梅查帮玲徐红梅
Owner 合肥欧创基因生物科技有限公司
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