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Pratylenchus penetrans detection kit and detection method thereof

A technology of short-bodied puncture nematodes and kits, which is applied in the application field of port inspection and quarantine laboratories, can solve the problems of long identification period, identification relying on experience, and inability to meet the rapid customs clearance of imported seedlings, etc., and achieve fast detection speed and specificity Strong, highly sensitive effects

Inactive Publication Date: 2015-04-22
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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AI Technical Summary

Problems solved by technology

[0003] At present, domestic detection methods for Brachyphyllo punctatus still rely on classical morphological methods, which are limited by the growth and development period and the number of worm populations, resulting in the identification of this nematode relying on experience, and there are deficiencies in the long identification period and other aspects. , cannot meet the needs of rapid customs clearance of imported seedling commodities

Method used

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  • Pratylenchus penetrans detection kit and detection method thereof
  • Pratylenchus penetrans detection kit and detection method thereof
  • Pratylenchus penetrans detection kit and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Embodiment 1, the detection of short-bodied nematode puncture

[0041] DNA extraction: According to the instructions of the TIANamp Genomic DNA Kit Tissue Genomic DNA Extraction Kit, the extracted nematode genomic DNA was dissolved in 50uL TE and stored at -20°C for later use.

[0042] PCR amplification: Sterile water was used as a blank control, and P.cof1, P.cof2, P.cof3, P.scr1, P.scr2, P.neg1, P.neg2, P.vul1, P.pen1, P.pen2, P.pen3 and P.pen4 nematode genomic DNA were used as templates for PCR amplification.

[0043] PCR reaction system:

[0044]

[0045] PCR reaction conditions: pre-denaturation at 94°C for 1 min, denaturation at 94°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 40 s, 35 cycles, and extension at 72°C for 5 min.

[0046] result:

[0047] from figure 1 It can be seen from the results that samples 9, 10, 11, and 12 are Brachybody punctures, all of which have a single band of 634 bp size specificity; The samples No. 6 and No. 7 w...

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Abstract

Relating to the quarantine inspection field of plant diseases and insect pests, the invention provides a Pratylenchus penetrans detection kit and a detection method thereof. By designing specific primers specific to the 18S specific sequence of Pratylenchus penetrans and applying a PCR amplification technology, rapid detection of Pratylenchus penetrans is realized. With the characteristics of strong specificity, high sensitivity and good stability, the method plays an important role in improving the accuracy of Pratylenchus penetrans detection and shortening the detection time.

Description

technical field [0001] The invention relates to the field of quarantine of plant diseases and insect pests, and provides a method for rapidly detecting Brachyphytes puncture nematodes by using PCR amplification technology and a kit thereof, which are suitable for application in port inspection and quarantine laboratories. Background technique [0002] Pratylenchus penetrans belongs to Pratylenchus family, Pratylenchus subfamily, and genus Pratylenchus. It has a wide range of hosts. There are more than 350 species of plant hosts reported. It mainly harms important economic crops such as wheat, corn, barley, potatoes, and lilies. Tulips, tulips and other flowers are currently my country's entry plant quarantine pests, and are one of the key inspection targets for my country's imported seed products. Brachybody puncture nematode is a migratory endoparasitic nematode, which is distributed all over the world. It has been reported in all temperate countries in the world, as well as...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 郭京泽张裕君王金成张瑞峰魏亚东廖芳
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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