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The Specific Primer Pairs and Their Application for Identifying Tyrophaphaphae mite and Acarina ellipsoides

A kind of technology of ellipse mealy mites and primer pairs, which can be used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. , strong specificity and high sensitivity

Active Publication Date: 2022-04-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the small size of Tyrophae saprophagae and Aurophagus ellipsoidus, slide specimens need to be prepared for morphological identification, and the identification mainly depends on the position and length of bristles, which has caused great difficulties for the morphological identification of the two species of mites

Method used

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  • The Specific Primer Pairs and Their Application for Identifying Tyrophaphaphae mite and Acarina ellipsoides
  • The Specific Primer Pairs and Their Application for Identifying Tyrophaphaphae mite and Acarina ellipsoides
  • The Specific Primer Pairs and Their Application for Identifying Tyrophaphaphae mite and Acarina ellipsoides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1, the identification of the acquisition of specific primer pairs for Tyrophaphae spp. and T. ellipsoides

[0096] 1. Mite mitochondrial genome acquisition

[0097] 1. Test mite samples

[0098] The tested Trichomonas ellipsoides were collected in the Czech Republic, and the tested Tyrophagous spp. were collected in Beijing, China. The samples were soaked in absolute ethanol and stored at -20°C for later use.

[0099] 2. Mitochondrial Genome Sequence

[0100] The micro DNA extraction kit from Tiangen Company was used to extract the DNA of the tested mite samples. The components of the micro DNA extraction kit are shown in Table 1.

[0101] Table 1. Components of Micro DNA Extraction Kit

[0102] Kit composition (50 times) The amount of each component Buffer GA 15ml Buffer GB 15ml Buffer GD 13ml Rinsing solution PW 15ml Elution buffer TB 15ml Proteinase K 1ml Carrier RNA 310μg RNase-free ddH 2 o

...

Embodiment 2

[0136] Example 2. Specific detection of specific primer pairs for identification of Tyrophagous putrescens and T. ellipsoides

[0137] 1. Specific detection method for specific primer pairs for identifying Tyrophagous putrescens and A. ellipsoides

[0138] 1. Test mite samples

[0139] The tested mites are shown in Table 5. The sample mites were all placed in absolute ethanol and stored at -20°C for later use.

[0140] Table 5. Identification of mite samples used in specific primer-specific PCR

[0141] Chinese name division Latin name ellipsoid mite Acaridae Aleuroglyphus ovatus A. rough-legged mite Acaridae Acarus siro Tyrophagous mite Acaridae Tyrophagus putrescentiae Tyrophilus linea Acaridae Tyroborus lini sweet mite Glycophidae Glycyphagus domesticus common carnivorous mite Carnivorous Acaridae Cheyletus eruditus Malacca carnivorous mite Carnivorous Acaridae Cheyletus malaccensis

[0142] ...

Embodiment 3

[0166] Example 3, Sensitivity detection of specific primer pairs for identification of Tyrophagous putrescens and T. ellipsoides

[0167] 1. Sensitivity detection method for specific primer pairs for identification of Tyrophaphae spp. and T. ellipsoides

[0168] Putty mite: use the same mite sample as in Example 1, dilute the DNA sample of putrefaction mite, and obtain concentrations of 26.62ng / ul, 13.31ng / ul, 6.66ng / ul, 3.33ng / ul and 1.66ng DNA samples per ul were amplified using the primer pair Tp-F / Tp-R using the diluted DNA samples as templates. The PCR amplification system and amplification conditions were the same as those in Step 2 of Example 1.

[0169] A. ellipsoides: use the same mite sample as in Example 1, dilute the DNA samples of A. ellipsoides to obtain concentrations of 15ng / ul, 5ng / ul, 1.67ng / ul, 0.56ng / ul and 0.18ng respectively The DNA samples / ul were amplified using the diluted DNA samples as templates, using the primer pair Ao-F / Ao-R, and the PCR amplific...

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Abstract

The invention discloses a pair of specific primers for identifying tyrophagous mite and T. ellipsoida and application thereof. The present invention is based on the conventional PCR molecular identification technology to realize the species identification of tyrophagous and adenophora ellipsoides, and the specific primer pair designed by the present invention for identifying tyrophagous tyrophagous and acanthus ellipsoids has strong specificity, high sensitivity and low cost , simple operation, and short time-consuming advantages, not only solves the problem of species identification of non-adult mites and mite adult residues, but also contributes to the epidemic monitoring, distribution and invasion status of tyrophagous mite and ellipsoid mite in my country, and Its subsequent governance provides practical technology and theoretical basis. It is of great significance to effectively prevent its further invasion and spread, protect agricultural production and ecological security, and promote the development of my country's economy and trade.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a pair of specific primers for identifying tyrophagous mite and acarina ellipsoidus and application thereof. Background technique [0002] Mites belong to the order Arachinida, Acari, and Acariformes. The individuals are tiny, and the adult mites are generally 0.1-0.2mm in length, which is difficult to observe with the naked eye. Storage pests can breed in large quantities in stored goods, eat the nutrient part of stored goods, and cause the loss of quality and quantity of stored goods. It will quickly cause large-scale pollution and mildew of the stored goods. More importantly, most species of storage pest mites can cause allergic diseases such as asthma and endanger people's health. [0003] According to the survey of stored goods in recent years, it is found that tyrophagous mite is the dominant species among storage pests, and the detection rate is the highe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686
Inventor 李志红蓝杨铭
Owner CHINA AGRI UNIV
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