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Method of synchronous detecting lily mottle virus, lily symptom less virus and cucumbus flomer leaf virus

A technology for simultaneous detection and virus detection, applied in biochemical equipment and methods, preparation of samples for testing, measurement devices, etc. Saving detection time and reagent costs, the effect of saving time and costs

Inactive Publication Date: 2006-06-28
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ELISA method and the ordinary RT-PCR method can only detect one virus at a time. For the case of multiple viruses co-infecting lilies, each virus needs to be detected separately, which wastes time and increases the detection cost. input
In addition, in the ordinary RT-PCR amplification process, upstream and downstream primers need to be designed separately for each virus, resulting in increased detection costs and increased the probability of non-specific detection

Method used

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  • Method of synchronous detecting lily mottle virus, lily symptom less virus and cucumbus flomer leaf virus

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0030] The known lily plants co-infected with three viruses of LMoV, LSV and CMV were used as materials, and healthy lily plants were used as negative controls, and this method was used for detection.

[0031] 1) Design primers:

[0032] (a) Design and detect the downstream general primer LMSV of two kinds of viruses of LMoV and LSV:

[0033] LMSV 5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTTTV-3 V = A, C or G

[0034] Primer name

Primer sequence

LMoV-F

LSV-F

5-AAGAAGCACGCTGGACTGC-3

5-ATCGGAGGCCGCTCTTCG-3

[0035] Primer name

Primer sequence

CMV-F

CMV-R

5-TCGTCCGCGTCGCGGTTC-3

5-TGGGAATCCCTTGGTGCTC-3

[0036] Primer name

Primer sequence

Amplified fragment size

Viruses detected

LMoV-F

LMSV

LSV-F

LMSV

CMV-F

CMV-R

5-AAGAAGCACGCTGGACTGC-3

5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTV-3

5-ATCGGAGGCCGCTCTTCG-3

5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTV-3...

example 2

[0053]The known lily plants infected with LMoV, LSV or CMV were tested, and healthy lily was used as negative control.

[0054] 1) Design primers:

[0055] (a) Design and detect the downstream general primer LMSV of two kinds of viruses of LMoV and LSV:

[0056] LMSV 5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTTTV-3 V = A, C or G

[0057] Primer name

Primer sequence

LMoV-F

LSV-F

5-AAGAAGCACGCTGGACTGC-3

5-ATCGGAGGCCGCTCTTCG-3

[0058] Primer name

Primer sequence

CMV-F

CMV-R

5-TCGTCCGCGTCGCGGTTC-3

5-TGGGAATCCCTTGGTGCTC-3

[0059] Primer name

Primer sequence

Amplified fragment size

Viruses detected

LMoV-F

LMSV

LSV-F

LMSV

CMV-F

CMV-R

5-AAGAAGCACGCTGGACTGC-3

5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTV-3

5-ATCGGAGGCCGCTCTTCG-3

5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTV-3

5-TCGTCCGCGTCGCGGTTC-3

5-TGGGAATCCCTTGGTGCTC-3

1153bp

...

example 3

[0076] The known lily plants co-infected with LMoV and LSV, the lily plants co-infected with LSV and CMV, and the lily plants co-infected with LMoV and CMV were used as materials, and healthy lily plants were used as negative controls, and the method was used for detection.

[0077] 1) Design primers:

[0078] (a) Design and detect the downstream general primer LMSV of two kinds of viruses of LMoV and LSV:

[0079] LMSV 5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTTTV-3 V = A, C or G

[0080] Primer name

Primer sequence

LMoV-F

LSV-F

5-AAGAAGCACGCTGGACTGC-3

5-ATCGGAGGCCGCTCTTCG-3

[0081] Primer name

Primer sequence

CMV-F

CMV-R

5-TCGTCCGCGTCGCGGTTC-3

5-TGGGAATCCCTTGGTGCTC-3

[0082] Primer name

Primer sequence

Amplified fragment size

Viruses detected

LMoV-F

LMSV

LSV-F

LMSV

CMV-F

CMV-R

5-AAGAAGCACGCTGGACTGC-3

5-GATGGCTGACTGGATCCTTTTTTTTTTTTTTTTV...

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Abstract

A method for synchronously detecting lily mottling virus, lily silent virus and cucumber mosaic virus includes carrying out amplification on three viruses of LMoV, LSV and CMV infected by lily in the same reaction tube and under the same heat circulation condition after detection processes of pick ¿C up total RNA, multiple RT ¿C PCR and electrophoresis; then carrying out separation and detection by means of gel electrophoresis.

Description

Technical field: [0001] The invention relates to a multiple RT-PCR method for synchronously detecting three main viruses of lily mottle virus (LMoV), lily asymptomatic virus (LSV) and cucumber mosaic virus (CMV) infecting lilies, belonging to the field of plant protection. Background technique: [0002] Lily is an important economic crop that integrates ornamental, edible, and medicinal uses. Especially the cut lily is bright in color, elegant in temperament, and has a unique fragrance. It has become one of the most important ornamental flowers in the world and has a high economic value. value. In recent years, with the continuous improvement of people's living standards, the demand for fresh cut flowers, especially lilies, has been increasing, and the planting scale of lilies has been expanding day by day. However, due to non-standard production methods and long-term asexual reproduction, viruses continue to accumulate, and the occurrence of viral diseases in lily is becom...

Claims

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Application Information

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IPC IPC(8): G01N21/64C12Q1/02G01N1/28G01N21/78G01N27/447
Inventor 李凡孙健陈海如李正跃王钰丽刘云龙范静华姬广海孔宝华王扬杨斌蒋小龙白松蔡红胡先奇
Owner YUNNAN AGRICULTURAL UNIVERSITY
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