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Method and kit for detecting multiple encephalitis related viruses

A virus and encephalitis technology, applied in the field of detection of various encephalitis-related viruses, can solve the problems of lack of encephalitis-related viruses, missed detection, etc.

Active Publication Date: 2012-11-28
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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  • Method and kit for detecting multiple encephalitis related viruses
  • Method and kit for detecting multiple encephalitis related viruses
  • Method and kit for detecting multiple encephalitis related viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Embodiment 1 Detection of encephalitis-related virus

[0112] 1. Coating of probe microspheres:

[0113] 1. Follow the list below to synthesize probes

[0114]

[0115] Note: The spacer arm is a 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 equal to poly(T) 10 +SEQ ID NO.: 7, and so on.

[0116] 2. Select 6 kinds of fluorescently encoded microspheres No. 24, 28, 31, 33, 36 and 42 [purchased from Luminex Company], and carry out the coating of the probes corresponding to the probes in the above table in turn. The methods are 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) Dissolve each corresponding probe with double distilled water, the concentration is 0.01mM (10pmol / μL);

[0119] (3) Mix the microspheres evenly with a shaker;

[0120] (4) Take 50ul (6.0×10 5 ) microspheres, put them into pre-marked cl...

Embodiment 2

[0187] Embodiment 2 Detection of encephalitis-related virus

[0188] 1. Coating of probe microspheres:

[0189] 1. Follow the list below to synthesize probes

[0190]

[0191] 2. Select 6 kinds of fluorescently encoded microspheres No. 24, 28, 31, 33, 36 and 42 [purchased from Luminex Company], and carry out the coating of the probes corresponding to the probes in the above table in turn. The methods are as follows:

[0192] (1) Equilibrate various microspheres and a portion of EDC powder stored at -20°C at room temperature for 30 minutes;

[0193] (2) Dissolve each corresponding probe with double distilled water, the concentration is 0.01mM (10pmol / μL);

[0194] (3) Mix the microspheres evenly with a shaker;

[0195] (4) Take 50ul (6.0×10 5 ) microspheres, put them into pre-marked clean 1.5ml centrifuge tubes;

[0196] (5) Centrifuge the microspheres for 5 minutes under the condition of 8000g centrifugal force, and carefully discard the supernatant;

[0197] (6) Add ...

Embodiment 3

[0258] Embodiment 3 Detection of encephalitis-related virus

[0259] Example 2 was repeated, except that samples 9-14 were replaced with samples 15 and 16. Among them, the No. 1 and No. 9 samples were combined as the No. 15 sample. Sample No. 4 and No. 10 were combined as sample No. 16.

[0260] The test results showed that Sample No. 15 contained Eastern Equine Encephalitis Virus and Japanese (Type B) 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 present invention relates to the technical fields of molecular biology and nucleic acid detection. Specifically, the present invention relates to methods and kits for the detection of various encephalitis-related viruses. Background technique [0002] Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), Venezuelan equine encephalitis virus (VEEV), Tick-borne encephalitis virus (VEEV) encephalitis virus, TBEV), Japanese encephalitis virus (Japanese encephalitis virus, JEV) are highly pathogenic viruses. [0003] Eastern equine encephalitis virus, western equine encephalitis virus and Venezuelan equine encephalitis virus are typical zoonotic viral diseases, which belong to the genus Alphavirus of the group A arbovirus family Togaviridae, and contain single-stranded RNA. The virus was successively isolated from the brain tissue of sick horses in the eastern, western and Venezuelan United States, hence the name. Subseq...

Claims

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Application Information

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Patent Type & Authority Applications(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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