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Zika virus fluorescent PCR detecting kit

A detection kit, Zika virus technology, applied in the direction of microbial determination/inspection, microorganisms, biochemical equipment and methods, etc., can solve the problem of no newborn baby, shorten the time required for detection, reduce workload, Guaranteed effect of specificity and accuracy

Inactive Publication Date: 2016-07-06
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Zika virus nucleic acid can be detected in breast milk, but infection of newborns through breastfeeding has not been reported

Method used

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  • Zika virus fluorescent PCR detecting kit
  • Zika virus fluorescent PCR detecting kit
  • Zika virus fluorescent PCR detecting kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 A kind of Zika virus fluorescent PCR detection kit and its use

[0023] 1. Prepare a kit comprising the following components:

[0024] 1 tube of primer-probe mixture (25 μl / tube), 1 tube of RT-PCR reaction solution (125 μl / tube), 1 tube of RT-PCR enzyme system (75 μl / tube), 1 tube of positive quality control (200 μl / tube), Negative quality control (200μl / tube) 1 tube, DEPCH 2 O (2000μl / tube) 1 tube.

[0025] 2. Preparation of target gene recombinant plasmid

[0026] The entrusted company synthesized and purified the in vitro transcribed RNA of Zika virus NS1 gene, measured the concentration by UV spectrophotometer, then calculated the RNA copy number, and used DEPCH 2 O were diluted to 1.0 × 10 10 copies / mL~1.0×10 1 copies / mL of the standard.

[0027] 3. Real-time fluorescent quantitative PCR amplification and detection

[0028] 3.1 Reagent preparation:

[0029] Take the corresponding amount of primer-probe mixture 1 μl, RT-PCR reaction solution 5 μ...

Embodiment 2

[0038] Example 2 Application of a Zika virus fluorescent PCR detection kit to detect specific samples

[0039] Select dengue virus type 1, dengue virus type 2, dengue virus type 3, dengue virus type 4, chikungunya virus, fever with thrombocytopenia syndrome virus, Hantaan virus, Seoul virus, Puumala virus , Japanese encephalitis virus, chikungunya virus, Epstein-Barr virus, human cytomegalovirus, and measles virus, each positive sample was used as a specific sample. Nucleic acid was extracted from all samples, and the steps of PCR amplification and result analysis were implemented by reference Example 1 is carried out, and the detection of negative and positive quality control products is carried out at the same time.

[0040] Test result: the amplification curve of the negative quality control product is not S-shaped (see attached image 3 ), the amplification curve of the positive quality control product is an obvious S-shaped curve (see attached Figure 4 ), both negative...

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Abstract

The invention provides a Zika virus fluorescent PCR detecting kit, and relates to a kit for detecting Zika virus, in particular to simultaneous detection of Zika virus RNA by using a real-time fluorescent polymerase chain type reaction technology.The kit mainly comprises RT-PCR reaction liquid, primer probe mixed liquid, an RT-PCR reaction enzyme system, DEPC H2O and a packaging box for separating and packaging reagent bottles or tubes in a centralized manner.The kit adopts a Taqman probe detection mode and a one-step real-time fluorescent PCR reaction mode, can accurately and quickly detect the Zika virus RNA, and can be widely applied to various fields such as Zika virus disease clinic early diagnosis, disease prevention and scientific research.

Description

technical field [0001] The invention relates to a Zika virus fluorescent PCR detection kit, in particular to a kit for detecting the Zika virus NS1 gene in one step using real-time fluorescent polymerase chain reaction technology. Background technique [0002] Zika virus disease (Zika Virus Disease) is a self-limiting acute disease caused by Zika virus (Zika Virus) and transmitted by mosquitoes, mainly through the bite of Aedes aegypti. The clinical features are mainly fever, rash, arthralgia or conjunctivitis, and rarely cause death. The World Health Organization (WHO) believes that neonatal microcephaly and Guillain-Barré syndrome may be related to Zika virus infection. The bite of infected Aedes mosquitoes is the main route of transmission of the disease. The main vector is Aedes aegypti, although Aedes albopictus, Aedes africanus and Aedes yellow-headed mosquitoes may also transmit the virus. It can also be transmitted from mother to child, including intrauterine infe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q1/701C12Q2563/107
Inventor 李明何皖平周其伟蒋析文高秀洁夏乔尤欢叶爱华吴洁
Owner DAAN GENE CO LTD
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