Fluorescent quantitative PCR detection method of resistance of bemisia tabaci on thiamethoxam and kit thereof

A technology of Bemisia tabaci and thiamethoxam, applied in the field of fluorescent quantitative PCR detection method and its kit, can solve the problems of low sensitivity, long cycle, poor repeatability, etc., and achieve the effect of strong specificity, high sensitivity and accuracy

Inactive Publication Date: 2014-09-10
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention aims at the problems of low sensitivity, long period, poor repeatability and high material requirements existing in the existing detection technology of Bemisia tabaci resistance to thiamethoxam. A fast and accurate fluorescent quantitative PCR molecular detection method and a fast, simple, accurate and efficient detection kit for the detection of thiamethoxam resistance in whitefly, providing an effective tool for the effective detection of thiamethoxam resistance in whitefly

Method used

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  • Fluorescent quantitative PCR detection method of resistance of bemisia tabaci on thiamethoxam and kit thereof
  • Fluorescent quantitative PCR detection method of resistance of bemisia tabaci on thiamethoxam and kit thereof
  • Fluorescent quantitative PCR detection method of resistance of bemisia tabaci on thiamethoxam and kit thereof

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Experimental program
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Effect test

Embodiment 1

[0045] The detection of TH-S and TH-R adult CDNA samples with fluorescence quantitative PCR technology and the establishment of the test kit and its usage method.

[0046] 1. Tobacco CYP4C64 The conservative segment area of the gene is the target target sequence (sequence 3 in the sequence list). According to the principle of the design principle of the fluorescent quantitative PCR primer, the design of specific fluorescent quantitative primers is as follows:

[0047] Positive primer (Q C64 -F): 5′-GCACACACCACACCCCGTAGCAG-3 ′ (sequence 1 in the serial list 1);

[0048] Reverse primer (Q C64 -R): 5′-GGCTCTTCGACACATCG-3 ′ (sequence 2 in the serial list);

[0049] This specific primer amplification fragment sequence figure 1 Show, its extension specificity is figure 2 (Among them, 1: CDNA samples sensitive to tosinine; 2: CDNA samples that are resistant to cycamine; M: Marker I), from the figure, we can see that the primer has high enlargement specificity and amplification, amplifi...

Embodiment 2

[0084] Three other three ticatizide resistant tobacco powder group ZJ, TJ, and BJ verification practices 1 of the kit and detection methods in the testing method 1 are shown in the specific experimental process.

[0085] Final fluorescent quantitative PCR reaction detection result is Figure 6 Among them, A (TH-R): Example 1 Smooth powder group TH-R adult CDNA-positive control samples; B (TH-S): Entrobutic 1 pairs in Example 1Timorizine-sensitive nicotin lice population TH-S adults CDNA negative control sample; C (ZJ): Tobacco powder lice furry group ZJ adult CDNA to be tested in Zhejiang: D (TJ): Tobaccum lice breeding group TJ adult CDNA to be tested in Tianjin area to be tested in Tianjin.Sample test.

[0086] Under specific wavelength conditions, the results of the CT value of the anti -allergic sample between fluorescent detection are judged, we can see that the two samples produce a typical amplification curve, and the CT value of the ZJ sample is 23.51 ± 0.19 (SE).The CT val...

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Abstract

The invention discloses a specific PCR (Polymerase Chain Reaction) primer pair for detecting the resistance of (i)Bemisia( / i)(i)tabaci( / i) on thiamethoxam, a kit and a detection method. The forward primer nucleotide sequence of the specific PCR primer pair is as shown in SEQ ID No.1, and the reverse primer nucleotide sequence of the specific PCR primer pair is as shown in SEQ ID No.2. The fluorescent quantitative PCR detection method of the bemisia tabaci on the thiamethoxam, which is disclosed by the invention, has the characteristics of less labour power, small sample quantity, large detection quantity, high sensitivity, high specificity, fastness, simpleness, convenience, accuracy and reliability, thereby effectively detecting the resistance of the bemisia tabaci on the thiamethoxam; the fluorescent quantitative PCR detection method disclosed by the invention is suitable for the early fast diagnosis of the resistance of the bemisia tabaci on the thiamethoxam, can be used for the real-time monitoring, prewarning and prediction of the resistance of the field bemisia tabaci on the thiamethoxam and is wide in application prospect.

Description

Technical field [0001] The invention is a biological field, which involves the fluorescent quantitative PCR detection method and their kits involved in nicotin lice. Background technique [0002] Nicheflyfront BEMISIA TABACI (Gennadius) is a worldwide agricultural pest, which has a great harm to production.After the 1980s, it was distributed in more than 90 countries and regions of the world's continents outside the Antarctica, and there were more than 74 families and 500 plants.In China, the pest became an important economic crop pest in my country in the late 1990s, mainly for the type of harm type B biological type. Because of its strong hazardity and harmfulness, it was also crowned as "super pests"; For the first time, this laboratory was discovered in the flower market in Yunnan, and then quickly spread to all parts of the country. At present, these two biological smoke powder are the most harmful biological types in Chinese vegetables and flowers.Tobacco mesh not only stab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 张友军杨鑫谢文王少丽吴青君李如美杨妮娜郭丽桃刘雅婷
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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