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Multiple PCR detection method for detecting symbiotic bacteria in tetranychus truncatus ehara body

A technology of symbiotic bacteria and mites, applied in the biological field, can solve the problems of consumables, labor consumption, and time consumption, and achieve the effects of reducing costs, improving efficiency, and saving time

Inactive Publication Date: 2009-12-16
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

When a large number of samples need to be tested, it is time-consuming, consumable and labor-intensive to test Wolbachia and Cardinium respectively.

Method used

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  • Multiple PCR detection method for detecting symbiotic bacteria in tetranychus truncatus ehara body
  • Multiple PCR detection method for detecting symbiotic bacteria in tetranychus truncatus ehara body
  • Multiple PCR detection method for detecting symbiotic bacteria in tetranychus truncatus ehara body

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Embodiment Construction

[0040] 1. Suitable for multiplex PCR (Multiplex PCR) specific primer design

[0041] The symbiotic bacteria Wolbachia pipientis, referred to as Wolbachia, is well known and commonly used, see references (Hertig M. The rickettsia, Wolbachia pipientis (gen. et sp.n.) and associated inclusions of the mosquite Culex pipiens. Parasitology, 1936, 28: 453-486 ), the Chinese genus name is Wolbachia, and there is no Chinese species name.

[0042]The symbiotic bacterium Cardinium, that is, Candidatus Cardinium hertigii, is well known and public, see references (Zchori-Fein E, Perlman S J, Kelly S E, Katzir N, Hunter M S. Characterization of a'Bacteroidetes' symbiont in Encarsia wasps (Hymenopera: Aphelinidae): proposal of'Candidatus Cardinium Hertigii', Int.J.Syst.Evol.Microbiol., 2004, 54:961-968.), first named in 2004, no Chinese generic name yet.

[0043] Wolbachia's epidermal protein gene wsp gene is often used as a genetic marker for the detection of Wolbachia. So far, only one g...

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Abstract

The invention relates to a multiple PCR detection method for detecting Wolbachia and Cardnium symbiotic bacterias in tetranychus truncatus ehara body at the same time, belonging to the field of agricultural biotechnologies; the multiple PCR is an improved PCR technology on the basis of a common PCR, a plurality of pairs of auele specific primers are added in a PCR reaction system, aiming at different areas of a plurality of DNA templates or the same template, a plurality of purpose fragments are amplified. According to Wolbachia wsp gene sequence and Cardnium 16S rDNA gene sequence, two pairs of new auele specific primers are designed, and two purpose fragments - 211bp and 279bp which has different size, are amplified. The multiple PCR detection method has high specificity, sensibility, feasibility and effectiveness, namely, the detecting cost is saved and the template is saved. The multiple PCR detection technology for detecting the Wolbachia and Cardnium symbiotic bacterias at the same time can be widely applied to the detection of the two symbiotic bacterias in tetranychus truncatus ehara.

Description

1. Technical field [0001] This study developed a multiplex PCR method that can simultaneously detect the symbiotic bacteria Wolbachia and Cardinium in the spider mite, which belongs to the field of biotechnology and can be used to detect these two symbiotic bacteria in the spider mite. 2. Background technology [0002] Wolbachia and Cardinium are two types of maternally inherited commensal bacteria that can regulate the reproduction of their hosts through different methods. Wolbachia, belonging to the α subphylum, can cause the following four reproductive abnormalities in the host: cytoplasmic incompatibility, parthenogenesis, feminization, andricide. Caidinium belongs to the Bacteroidetes bacterial group and can cause three abnormal reproductive behaviors in the host: cytoplasmic incompatibility, parthenogenesis, and feminization. [0003] With the deepening of the research, it was found that in the reproductive system of a few insects and mites, there are two types of par...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/01
Inventor 洪晓月陈小琳谢蓉蓉
Owner NANJING AGRICULTURAL UNIVERSITY
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