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PCR kit for testing banana apical tuft virus and its test method

A technology of banana bunchy top virus and detection method, which is applied in biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problems of easy false positive reaction, high technology and equipment conditions, low detection sensitivity, etc., and achieve improvement The effect of virus-free detection level, cost reduction, and simple detection

Inactive Publication Date: 2004-06-23
福建省农业科学院生物技术中心
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the detection of banana bunchy top virus (abbreviated as BBTV) is mainly based on enzyme-linked immunosorbent assay (abbreviated ELISA), which has the following disadvantages: false positive reactions are prone to occur during the detection process; the detection effect on diseased strains with low virus content Not ideal, that is, the detection sensitivity is low; more importantly, as an indispensable reagent for the ELISA method, the preparation process of the monoclonal antibody against banana bunchy top virus is complicated, requires high technology and equipment conditions, and is difficult to promote and use on a large scale

Method used

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  • PCR kit for testing banana apical tuft virus and its test method
  • PCR kit for testing banana apical tuft virus and its test method
  • PCR kit for testing banana apical tuft virus and its test method

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Embodiment Construction

[0058] The present invention will be described in more detail below in conjunction with examples.

[0059] Best practice:

[0060] 1. Preparation of materials and equipment required for testing experiments

[0061] 1. Kit composition:

[0062] 1) MgCl 2 (25mM) 50μl

[0063] 2) dNTP (2mM) 50μl

[0064] 3) 10×buffer (10×enzyme-specific reaction buffer) 100μl

[0065] 4) Taq polymerase (3U / μl) (Taq polymerase) 10μl

[0066] 5) Primer mixture (primer) (50pmol) 30μl

[0067] 6) Banana bunchy top virus positive control (CK + ) 20μl

[0068] 7) Banana bunchy top virus negative control (CK - ) 20μl

[0069] 8)ddH 2 O 1ml

[0070] Each kit can be used to detect 20 samples.

[0071] where MgCl 2 , 10×buffer, and Taq polymerase were provided by Huamei Bioengineering Company; dNTP was provided by Shanghai Sangon Bioengineering Technology Service Co., Ltd.; Primermixture was provided by Shanghai Sangon Bioengineering Technology Service Co., Ltd. based on the sequence provided...

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Abstract

The present invention relates to microbe detecting composition and method, and is especially PCR kit and test method of testing banana apical tuft virus. The PCR test kit includes primer, dNTPs, buffering solution and DNA polymerase, and its upstream primer is 5-CTC GTC ATG TGC AAG GTT ATG TCG-3' and downstream primer is 5'-GAA GTT CTC CAG CTA TTC ATC GCC-3'. The PCR test kit is easy to produce and the test method of testing banana apical tuft virus has high sensitivity and high repeatability and is simple and practical. In detecting coarsely purified virus liquid, the method can detect banana apical tuft virus equivalent to the amount of 1 ng of banana tissue in high test precision and relatively low test cost.

Description

technical field [0001] The present invention relates to a kind of composition used in the measurement or inspection of microorganisms and a detection method thereof, in particular to a PCR kit which can be used to detect banana bunchy top virus and how to use the kit to detect banana bunchy top virus. Background technique [0002] Banana bunchy top disease is a common disease in the banana planting process. Its field incidence rate is generally 3-5%, and it can reach 30-50% in serious cases, even reaching more than 80%, seriously affecting the yield of bananas. [0003] In order to reduce this disease, it is generally necessary to detect the virus earlier when cultivating banana test-tube seedlings, then detoxify the toxic seedlings, and finally multiply the virus-free test-tube seedlings. [0004] At present, the detection of banana bunchy top virus (abbreviated as BBTV) is mainly based on enzyme-linked immunosorbent assay (abbreviated ELISA), which has the following disadv...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/25
Inventor 周莉娟郑伟文
Owner 福建省农业科学院生物技术中心
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