Multiplex real-time fluorescent polymerase chain reaction (PCR) primer for simultaneously detecting mouse adenovirus and poxvirus muris mouse pox virus, probe as well as reagent kit thereof

A mouse adenovirus and real-time fluorescence technology, which is applied in the field of multiple real-time fluorescent PCR primers and probes, can solve the problems of short detection time and achieve the effects of simple and fast operation, improved sensitivity and good linear relationship

Active Publication Date: 2012-04-04
STAIDSON (BEIJING) BIOPHARMACEUTICALS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve the deficiencies of existing mouse virus detection methods, the invention provides a multiple real-time fluorescent PCR primers, probes

Method used

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  • Multiplex real-time fluorescent polymerase chain reaction (PCR) primer for simultaneously detecting mouse adenovirus and poxvirus muris mouse pox virus, probe as well as reagent kit thereof
  • Multiplex real-time fluorescent polymerase chain reaction (PCR) primer for simultaneously detecting mouse adenovirus and poxvirus muris mouse pox virus, probe as well as reagent kit thereof
  • Multiplex real-time fluorescent polymerase chain reaction (PCR) primer for simultaneously detecting mouse adenovirus and poxvirus muris mouse pox virus, probe as well as reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Simultaneous detection of multiple real-time fluorescent PCR primers and probes for mouse adenovirus and decapitation virus

[0056] The full-length genome sequences of MADV-1, MADV-2, and MADV-3 were found from genbank, and their genbank numbers are NC_000942 / AC_000012, NC_014899 / H_049560, and NC_012584 / EU835513, respectively. Perform sequence comparison to find out the homologous sequence SEQ ID NO: 11 with its conserved sequence region;

[0057] According to the literature (David J.Esteban et al., 2005), the genome of the mouse deafness virus ECTV-Mos was analyzed, and combined with the literature (Gloria Ribas et al., 2003), the conserved gene EVM170 of ECTV-Mos was selected (ie crmD gene) for ECTV homology analysis. The gene sequence was compared with the GenBank+EMBL+DDBJ+PDB database, and with the mouse genome database to determine the homologous sequence SEQ ID NO:12.

[0058] Special primers and probe sequences for real-time fluorescent PCR detectio...

Embodiment 2

[0066] Example 2 Multiplex real-time fluorescent PCR detection kit and detection method for simultaneous detection of mouse adenovirus and decapitation virus

[0067] 2.1 Multiplex real-time fluorescent PCR detection kit for simultaneous detection of mouse adenovirus and decapitation virus

[0068] The components of the kit are:

[0069] Primer-probe mixture for two viruses (MADV reaction solution, EV reaction solution)

[0070] 2×Taqman PCR Mix

[0071] sterile ddH 2 o

[0072] Among them, 2×Taqman PCR Mix can directly use Taqman GeneExpression Master Mix, a product of AB Company.

[0073] Above-mentioned MADV reaction liquid is made up of special primers, probes that specifically amplify mouse adenovirus; EV reaction liquid is made up of special primers, probes that specificity amplifies mouse off-leg disease virus, wherein,

[0074] The volume ratio of MADV reaction solution to EV reaction solution is 1:1.

[0075] Wherein, the MADV reaction solution and the EV reacti...

Embodiment 3

[0117] Example 3 Multiple real-time fluorescent PCR detection kit and detection method for simultaneous detection of mouse adenovirus and decapitation virus

[0118] 3.1 Multiplex real-time fluorescent PCR detection kit for simultaneous detection of mouse adenovirus and foot disease virus

[0119] The components of this test kit differ from the components of the test kit in Example 2.1 only in that:

[0120] In the primer-probe mixture of the two viruses, the probe-labeled fluorescent groups of the two viruses are different, so that the kit can simultaneously detect the two viruses in a single tube.

[0121] Among them, the 5' end of the internal standard probe is marked with VIC, and the 3' end is marked with TAMRA; the detection probe of mouse adenovirus is marked with FAM at the 5' end, and the 3' end is marked with TAMRA; The 5' end of the probe was labeled with JOE, and the 3' end was labeled with TAMRA.

[0122] 3.2 Detection method of multiplex real-time fluorescent P...

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Abstract

The invention discloses a multiplex real-time fluorescent polymerase chain reaction (PCR) primer for simultaneously detecting mouse adenovirus and poxvirus muris mouse pox virus and a probe, which respectively contain oligonucleotides shown by SEQ ID NO: 1 to 6 in a sequence table. A multiplex real-time detection reagent kit containing the primer and the probe can be used for one-step combined detection of the mouse adenovirus and the poxvirus muris mouse pox virus, has the advantages of high speed, simplicity, high sensitivity, high specificity and high recovery rate, solves the problems of complexity, time waste, long period and certain requirement on an operation laboratory of the existing detection measure and has good application prospects.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a multiple real-time fluorescent PCR primer, a probe, a kit and a detection method for simultaneously detecting mouse adenovirus and alopecia areata virus. Background technique [0002] There are many viruses that naturally infect experimental animals. According to their hazards to humans, they can be divided into three categories; one category is zoonotic viruses, which can infect humans and primates; the second category has no signs of infecting humans. , but can replicate in human, ape and monkey-derived cells cultured in vitro, which is potentially dangerous to humans; the three types of viruses only infect animals themselves under natural conditions, and there is no sign that they can infect humans, so they are harmful to humans. Not much of a threat. [0003] Nowadays, as a major source of biological products such as monoclonal antibodies and protein drugs, mice have po...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 谭淑萍李萃王刚蒋立新周志文
Owner STAIDSON (BEIJING) BIOPHARMACEUTICALS CO LTD
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