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Parasite detection kit and detection method thereof

A technology for detection kits and detection methods, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc.

Inactive Publication Date: 2018-09-28
杭州泰熙生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The present invention aims at the disadvantages of requiring expensive equipment in the prior art, and provides a parasite detection kit and a detection method thereof

Method used

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  • Parasite detection kit and detection method thereof
  • Parasite detection kit and detection method thereof
  • Parasite detection kit and detection method thereof

Examples

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Embodiment 1

[0048] A parasite detection kit, such as Figure 1-5 As shown, amplification primers or RPA primers specifically amplify specific parasites to generate parasite-specific amplicons. Probes and amplicons are specific for amplifying Leishmania donovani, Leishmania chagasi, and Leishmania infantum. These parasite-specific amplicons were detected with oligonucleotide probes. Probes are specifically designed to target unique DNA sequences and are amplified by two primers. The combination of primers and probes determines the specificity (the DNA sequence to be detected) for the test. In certain aspects, the Leishmania donovani, Leishmania chagasi, Leishmania infantum specific amplification primer comprises an oligonucleotide sequence having 90%, 95%, 98% or 100% identity, an oligonucleotide The nucleotide sequence includes SEQ ID NO: 3, 4, 5, 6, 7, 8, 10, 11, 12, or 13. Amplification primers include: 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, ...

Embodiment 2

[0051]Primers, probes and amplicons are specific for the amplification of Leishmania brasiliensis. Oligonucleotide probes were used to detect these parasite-specific amplicons. The Leishmania brasiliensis amplification primers include 70, 75, 80, 85, 90, 95, 98 or 100% identical oligonucleotide sequences, and the oligonucleotide sequences include SEQ ID NO: 16, 17, 18, 19, 20, 21. In certain aspects, amplification primers include: , 45 consecutive oligonucleotides. Certain aspects relate to amplification primer pairs comprising SEQ ID NO: 16 / 19, 16 / 20, 16 / 21, 17 / 19, 17 / 20, 17 / 21, 18 / 19, 18 / 20, or 18 / 21. The oligonucleotide of the nucleotide sequence included in the RPA probe is SEQ ID NO:77.

Embodiment 3

[0053] Primers, probes and amplicons are specific for the amplification of the trypanosoma trypanosoma trypanosoma or trypanosoma rangeli. Oligonucleotide probes were used to detect these parasite-specific amplicons. In certain aspects, the Trypanosoma cruzi amplification primers comprise 90, 95, 98 or 100% identical oligonucleotide sequences comprising SEQ ID NO: 23, 24, 25, 26, 27, 28 . Amplification primers include: having 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 consecutive Oligonucleotides. Certain aspects relate to amplification primer pairs comprising SEQ ID NO: 23 / 26, 23 / 27, 23 / 28, 24 / 26, 24 / 27, 24 / 28, 25 / 26, 25 / 27, or 25 / 28. The RPA probe may comprise an oligonucleotide having the nucleotide sequence of SEQ ID NO: T. rangeli trypanosoma or SEQ ID NO: 29,30.

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Abstract

The invention relates to the field of parasite detection, and discloses a parasite detection kit and a detection method thereof. A nucleic acid reaction system of the parasite detection kit internallycomprises one or more nucleic acid amplification primer pairs for amplifying nucleotide sequences of DNA of trypanosome, leishmania, leishmania braziliensis, chagasi leishmania, Trypanoma cruzi, Trypanoma rangeli, leishmania, leishmania mexicana or leishmania amazonensis, panamensis leishmania donovani, guyanensis leishmania, or peruviana leishmania. The detection method comprises the steps thatthe one or more nucleic acid amplification primer pairs are used for a nucleic acid amplification reaction, and existence of the DNA of parasites is detected by detecting specific amplicons of the nucleic acid amplification primer pairs. Compared with a PCR, detecting through a RPA amplification mode is conducted in constant-temperature and simple equipment, the cost is low, the sensitivity is high, the parasite detection kit can be used for diagnosis of the trypanosome and the leishmania for early-infected people and infected dogs, and thus the VL incidence rate in human bodies is decreased.

Description

technical field [0001] The invention relates to the field of parasite detection, in particular to a parasite detection kit and a detection method thereof. Background technique [0002] Leishmaniasis, a disease of significant global impact affecting more than 88 countries in the world, is caused by the protozoa Leishmania. Cutaneous leishmaniasis (CL) is characterized by chronic skin ulceration that can affect an individual's functional status, lead to costly and untimely treatment, and lead to disfiguring scarring. Overall, there are 100,000-200,000 cases of leishmaniasis worldwide. Leishmaniasis results in a spectrum of oropharyngeal lesions including localized cutaneous leishmaniasis (LCL), destructive nasal and mucosal leishmaniasis (ML). LCL is the most common Viannia subgenus species (L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, L. (V.) peruviana) and is caused by Leishmania subgenus Genus and species are less severe (L.(V.) Mexico, L.(V.) Amazon). ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6893C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/6893C12Q2521/507C12Q2522/101Y02A50/30
Inventor 周斌王树明
Owner 杭州泰熙生物技术有限公司
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