Reverse transcription loop-mediated isothermal detection method for tomato chlorosis virus
The invention discloses a reverse transcription loop-mediated isothermal detection method for a tomato chlorosis virus. The invention discloses a group of primers, which is composed of DNA molecules represented by (1)-(4): (1) a DNA molecule represented by SEQ ID No.1; (2) a DNA molecule represented by SEQ ID No.2; (3) a DNA molecule represented by SEQ ID No.3; and (4) a DNA molecule represented by SEQ ID No.4. The invention provides a rapid, sensitive and high-specific method for detecting the tomato chlorosis virus, and provides new means for rapidly detecting the virus. 1. A group of primer, is composed of the following (1)-(4) shown by a DNA molecule: (1) ID SEQ No. 1 the DNA molecule is shown; (2) ID SEQ No. 2 the DNA molecule is shown; (3) ID SEQ No. 3 of the DNA molecule is shown; (4) ID SEQ No. 4 of the shown in the DNA molecule. 2. A detection tamarillo fade green virus of the kit, the kit comprises a primer according to Claim 1, reverse transcriptase and Bst DNA polymerase; Wherein the reverse transcriptase is M-MLV reverse transcriptase or AMV reverse transcriptase. 3. A detection tamarillo fade green method of virus, the method is to the total of the sample to be measured as the template RNA, to primer is primer for reverse transcription according to Claim 1 loop-mediated isothermal nucleic acid amplification, reverse transcription of loop-mediated isothermal nucleic acid amplification product, the candidate in said sample solution containing tomato green virus, if not to the reverse transcription loop-mediated isothermal nucleic acid amplification product, the candidate in said sample solution does not contain tomato green virus. 4. Method according to Claim 3, characterized in that the inverse transcription loop-mediated isothermal nucleic acid amplification method for judging whether or not the product is as follows (1) and/or (2): (1) the reverse transcription loop-mediated isothermal nucleic acid amplification reaction product to agarose gel electrophoresis, with braces nucleic acid electrophoresis strip of the sample to be measured containing tomato fade green virus sample, does not have the braces nucleic acid electrophoresis strip of the sample to be measured does not contain tomato fade green virus sample; (2) the reverse transcription loop-mediated isothermal nucleic acid amplification reaction product I with SYBR Green reacted reaction solution, the reaction to the sample to be measured of an emerald green for maching in order to contain the tomato to deliver green virus sample, reaction maching the orange of candidate sample to be measured does not contain tomato fade green virus sample. 5. Method as in Claim 3 or Claim 4, characterized in that the primers No. 1 ID SEQ shown in ID DNA molecules and SEQ No. 2 DNA molecule is shown in external primer according to Claim 1, SEQ No. 3 ID shown in ID DNA molecules and SEQ No. 4 DNA molecule is shown in the of the primer; SEQ No. 1 ID of the external composition of the DNA shown in ID SEQ No. 2 molecular DNA shown in the reverse transcription loop-mediated isothermal nucleic acid amplification in the molar concentration ratio of 1:1; The inner primer SEQ No. 3 ID in the ID of the DNA molecules SEQ No. 4 DNA molecules shown in the reverse transcription loop-mediated isothermal nucleic acid amplification in the molar concentration ratio of 1:1. The outer primer and the primer in the reverse transcription loop-mediated isothermal nucleic acid amplification in the molar concentration ratio of 1:4. 6. Any method according to Claim 3-5, characterized in that the inverse transcription loop-mediated isothermal nucleic acid amplification reaction temperature is in the 60 [...]. 7. Any method according to Claim 3-6, characterized in that the inverse transcription loop-mediated isothermal nucleic acid amplification in the concentration of the reverse transcriptase of 8U/the L; Reverse transcriptase of the trade names in particular to M-MLV reverse transcriptase, purchased from the Company TaKaRa, products for a directory 2641A. 8. Any method according to Claim 3-7, characterized in that the inverse transcription loop-mediated isothermal nucleic acid amplification of the system is as follows: 10 ×Thermopol buffer2.5 L, SEQ No. 1 ID of the DNA molecule in the 0.2 M, ID SEQ No. 2 molecular DNA shown in the 0.2 M, ID SEQ No. 3 of the DNA molecule in the 0.8 M, ID SEQ No. 4 DNA molecules shown in 0.8 M, dNTPs2.5mM, MgCl2 4 mm, reverse transcriptase 200U, Bst DNA polymerase 12U, RNA40ng the total of the sample to be measured, the rest is ddH2 O, system the 25 L; The buffer 10 ×Thermopol Newmont biotechnology England, purchased from (Beijing) limited, to a directory of products M0275; Wherein the reverse transcriptase is M-MLV reverse transcriptase or AMV reverse transcriptase; The DNA polymerase Bst Bst DNApolymerase trade names referred to, biological technology from newe England (Beijing) limited, to a directory of products M0275; The inverse transcription loop-mediated isothermal nucleic acid amplification reaction condition is:60 the thermostatic [...] 1h; the 80 [...] 5 min. 9. Any method according to Claim 3-8, characterized in that the reverse transcriptase M-MLV TaKaRa Company purchased, products for a directory 2641A; The sample in particular to a blade of the sample to be measured. 10. Claim 1 the primer in detecting tomato modifying the application of the green virus; And/or, Claim 2 the tomato detecting the reagent kit of the application of the green virus in solution.