Kit for detecting mosquito borne pathogens and detection method thereof

A detection method and pathogen technology, which are applied in the field of kits for detecting various mosquito-borne pathogens, can solve the problems of early diagnosis of unfavorable diseases, high labor and cost, and inability to meet the needs of critically infected patients.

Inactive Publication Date: 2014-07-09
河北国际旅行卫生保健中心
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  • Abstract
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Problems solved by technology

However, there are more or less defects in these methods: for example, the isolation and culture method is technically difficult, and it usually takes more than 3 to 5 days to obtain the results of etiological diagnosis and drug susceptibility, and it is difficult to carry out etiological diagnosis and drug resistance early in clinical practice. However, it cannot meet the needs of clinical treatment of critically ill infected patients; when immunological methods are used to detect virus-specific antibodies, it is not only necessary to collect double sera in the acute phase and

Method used

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  • Kit for detecting mosquito borne pathogens and detection method thereof
  • Kit for detecting mosquito borne pathogens and detection method thereof

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Embodiment Construction

[0025] 1. Synthesize the required primer pairs and probes according to the nucleic acid sequences in Table 1 and Table 2, and carry out biotin labeling on the 5' end of the downstream primer in each pair of primers; and carry out amine labeling on the 5' end of each probe Modified with a C12 contiguous arm sequence.

[0026] 2. Coupling the probes with corresponding fluorescently coded microspheres respectively, and then mixing them in equal numbers to prepare a microsphere group coupled with specific nucleic acid probes.

[0027] Select 12 types of microspheres (BioRad) numbered 22, 39, 15, 12, 18, 30, 36, 42, 45, 49, 56, and 52, and perform coupling of nucleic acid probes as follows:

[0028] 1. Store 12 microspheres numbered 22, 39, 15, 12, 18, 30, 36, 42, 45, 49, 56, 52 stored at 2-10°C and 2 bottles of unopened EDC stored at -20°C The powder (10mg / bottle) was placed at room temperature for 30-60 minutes to equilibrate.

[0029] 2. YFV-PF, CHIKV-PF, WNV-PF, JEV-PF, DV1...

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Abstract

The invention discloses a kit for detecting a plurality of mosquito borne pathogens and a detection method thereof. The plurality of mosquito-borne infectious pathogens are detected by using multiplex polymerase chain reaction (PCR) combined with a liquid chip, and a yellow fever virus (YFV), dengue fever viruses (DV) I-IV type, epidemic encephalitis B (japanese encephalitis) virus (JEV), plasmodium (Plasmodium), (including plasmodium vivax (PV), plasmodium falciparum (PF), plasmodium malariae (PM), plasmodium ovale (PO)), a west nile virus (MNV) and a chikungunya virus (CHIKV) can be detected in parallel one time. The kit has the advantages of being large in flux, high in specificity and sensitivity, stable in result, good in repeatability, simple to operate and fast in detection speed.

Description

technical field [0001] The invention belongs to the technical field of biochips and diagnostic reagents, in particular to a kit and a detection method for detecting various mosquito-borne pathogens, which design specific nucleic acid probes for various mosquito-borne infectious disease pathogens and react with multiple polymerase chain reactions Combining (PCR) technology and using liquid-phase chips to quickly and synchronously detect a variety of mosquito-borne pathogens and their typing. Background technique [0002] Mosquito-borne infectious diseases refer to diseases transmitted by mosquitoes biting the human body, and have been on the rise year by year in recent years. Once a mosquito-borne infectious disease becomes popular or breaks out, it will have a great impact on social stability, people's health, life and production. According to the needs of infectious disease screening at border ports, yellow fever, dengue fever, Japanese encephalitis (Japanese encephalitis)...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/04
CPCC12Q1/686C12Q1/70C12Q2531/113C12Q2563/149C12Q2563/131Y02A50/30
Inventor 史玲莉闫冀焕李云滑娜沈军吴志茹兰景李薇付晓昀
Owner 河北国际旅行卫生保健中心
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