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A method and kit for detecting multiple encephalitis-associated viruses

A technology of encephalitis virus and kit, which is applied in the field of molecular biology and nucleic acid detection, and can solve problems such as missed detection and lack of encephalitis-related viruses

Active Publication Date: 2016-11-09
SHANGHAI TELLGEN LIFE SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both methods have the disadvantage that only one virus can be detected at a time
Since the symptoms caused by several encephalitis viruses are relatively similar, only one virus (currently only Japanese encephalitis is often detected) will cause missed detection
[0009] In summary, there is still a lack of satisfactory techniques in the art that can simultaneously detect multiple encephalitis-associated viruses

Method used

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  • A method and kit for detecting multiple encephalitis-associated viruses
  • A method and kit for detecting multiple encephalitis-associated viruses
  • A method and kit for detecting multiple encephalitis-associated viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] The detection of embodiment 1 encephalitis-associated virus

[0112] 1. Coating of probe microspheres:

[0113] 1. Synthesize probes according to the list below

[0114]

[0115] Note: The spacer arm is poly(T) composed of 10 Ts 10 , so SEQ ID NO.: 12 is equal to poly(T) 10 +SEQ ID NO.: 6, SEQ ID NO.: 13 is equal to poly(T) 10 +SEQ ID NO.: 7, and so on.

[0116] 2. Select 6 kinds of fluorescently encoded microspheres with numbers 24, 28, 31, 33, 36 and 42 [purchased from Luminex], and coat the probes corresponding to the probes in the above table in turn. The method is as follows:

[0117] (1) Equilibrate various microspheres and a portion of EDC powder stored at -20°C at room temperature for 30 minutes;

[0118] (2) Take each corresponding probe and dissolve it in double distilled water, the concentration is 0.01mM (10pmol / μL);

[0119] (3) Mix the microspheres evenly with an oscillator;

[0120] (4) Take 50ul each (6.0×10 5 ) Microspheres, put into a clean ...

Embodiment 2

[0186] The detection of embodiment 2 encephalitis-associated virus

[0187] 1. Coating of probe microspheres:

[0188] 1. Synthesize probes according to the list below

[0189]

[0190] 2. Select 6 kinds of fluorescently encoded microspheres with numbers 24, 28, 31, 33, 36 and 42 [purchased from Luminex], and coat the probes corresponding to the probes in the above table in turn. The method is as follows:

[0191] (1) Equilibrate various microspheres and a portion of EDC powder stored at -20°C at room temperature for 30 minutes;

[0192] (2) Take each corresponding probe and dissolve it in double distilled water, the concentration is 0.01mM (10pmol / μL);

[0193] (3) Mix the microspheres evenly with an oscillator;

[0194] (4) Take 50ul each (6.0×10 5 ) Microspheres, put into a clean 1.5ml centrifuge tube with pre-marked numbers;

[0195] (5) Centrifuge under the condition of 8000g centrifugal force for 5 minutes to precipitate the microspheres, carefully discard the su...

Embodiment 3

[0257] The detection of embodiment 3 encephalitis-associated virus

[0258] Example 2 was repeated except that Samples 9-14 were replaced with Samples 15 and 16. Among them, samples No. 1 and No. 9 were combined as sample No. 15. Combine samples No. 4 and No. 10 as sample No. 16.

[0259] The test results showed that sample No. 15 contained Eastern equine encephalitis virus and Japanese encephalitis virus; sample No. 16 contained Venezuelan equine encephalitis virus and Western equine encephalitis virus. The test results are consistent with the actual situation of the samples.

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Abstract

The invention provides a method and a kit for detecting multiple encephalitis related viruses. In particular, the invention discloses a method for simultaneously detecting multiple encephalitis related viruses. The method comprises the following steps of: performing polymerase chain reaction (PCR) by using a specific primer set aiming at the encephalitis related viruses in a polymerase reaction system to obtain an amplification product; and detecting with specific probes or probe microspheres. The invention also provides the corresponding kit. The method and the kit can be used for sensitively and simply detecting and identifying multiple encephalitis related viruses comprising eastern equine encephalitis viruses, western equine encephalitis viruses, Venezuelan equine encephalitis viruses, forest encephalitis viruses and Japanese encephalitis viruses.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and nucleic acid detection. Specifically, the present invention relates to methods and kits for detecting various encephalitis-associated viruses. Background technique [0002] Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), Venezuelan equine encephalitis virus (VEEV), Tick-borne encephalitis virus (Tick-borne encephalitis virus) virus, TBEV), Japanese encephalitis (Japanese encephalitis) virus (Japanese encephalitis virus, JEV) are all highly pathogenic venereal disease viruses. [0003] Eastern equine encephalitis virus, western equine encephalitis virus and Venezuelan equine encephalitis virus are typical zoonotic viral diseases. The virus was isolated from the brain tissue of sick horses in the eastern and western parts of the United States and Venezuela, hence the name. Subsequently, the same virus was also isolated from the brain tissue of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 谭畅王勤熙张秀斐姚见儿
Owner SHANGHAI TELLGEN LIFE SCI CO LTD
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