CN100405061C - Pear virus hybridization detection kit and detection method thereof - Google Patents
Pear virus hybridization detection kit and detection method thereof Download PDFInfo
- Publication number
- CN100405061C CN100405061C CNB2005100653066A CN200510065306A CN100405061C CN 100405061 C CN100405061 C CN 100405061C CN B2005100653066 A CNB2005100653066 A CN B2005100653066A CN 200510065306 A CN200510065306 A CN 200510065306A CN 100405061 C CN100405061 C CN 100405061C
- Authority
- CN
- China
- Prior art keywords
- liquid
- virus
- sequence
- minutes
- pear
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
The present invention relates to a kit which is used for pear tree virus hybridization detection and a detecting method. The present invention is used for detecting three viruses of pear ring floral leaves, apple micropteres and xanthous vessels of pears on pear trees, the kit comprises a pre-hybridizing liquid, a hybridizing liquid, cleaning solutions I/II, buffer liquids I/II/III, an antigen liquid, primer sequence and probe sequence. The kit has the advantages of simple and convenient preparing method and easy industrialization production. The hybridization detecting method which is provided by the kit has the advantages of high stability, high sensitivity, strong specificity, quickness and simpleness, lower cost, etc. The present invention can be effectively applied to investigation and no virus standardization of pear virus types and distribution condition.
Description
Technical field
The present invention relates to a kind of pear virus hybridization detection kit and detection method thereof.
Background technology
The classic method of detection and evaluation fruit tree virus is indication plant method, serological method.Through screening decades, confirmed complete standard phyto-indicator in the world.This method is concerning identifying at present known fruit tree virus, though the reliability height, required time is longer, and the consumption ground of taking a lot of work.Serological method improves greatly than sensitivity and the detection speed that the phyto-indicator method detects fruit tree virus, and specificity strengthens, and has become conventional fruit tree virus at present and has detected and authentication method, and the research of relevant this respect has a lot of reports both at home and abroad.But this detection method causes because of the Antiserum Preparation of fruit tree virus difficulty and detects the cost height.And a lot of fruit tree virus can't use this method because of failing to prepare antiserum.
Since the eighties, the develop rapidly of Protocols in Molecular Biology is for the research of fruit tree virus provides a kind of unprecedented new way.Compared with phyto-indicator method and serological method, Protocols in Molecular Biology detects fruit tree virus its special advantages, the sensitivity of this technology for detection virus exceeds several magnitude than said method, can detect with said method detect less than virus, and specificity is stronger, speed is faster.More American-European countries take the lead in adopting Protocols in Molecular Biology to detect fruit tree virus.China starts late to the research of fruit tree virus disease.The development of plant virus detection kit seldom only has tobacco mosaic virus (TMV) detection kit etc. several.The Potyvirus detection kit has been developed in international potato research centre, and practical application.This kit has adopted serological detection method, can detect PVY, PVX, PVS, four kinds of viruses such as PLRV.A situation arises that huge impetus has been played in research to China's Potyvirus in the appearance of this kit.Along with the maturation of Protocols in Molecular Biology, a lot of methods have been applied to during fruit tree virus detects, but do not have about the research of fruit tree virus molecular detection kit.
Summary of the invention
The object of the invention is to provide a kind of pear virus hybridization detection kit and detection method thereof.
Content of the present invention is;
Pear virus hybridization detection kit and detection method thereof is characterized in that comprising:
The pear ring mosais virus probe sequence is as the sequence in the sequence table＜210〉shown in 1.
The apple stem grooving virus probe sequence is as the sequence in the sequence table＜210〉shown in 2.
The pear vein yellow virus probe sequence is as the sequence in the sequence table＜210〉shown in 3.
The amount of above-mentioned probe is: 0.005-0.03mg/mL DNA;
1# liquid, prehybridization solution contains 50% deionized formamide, 5 * Denhardt ' s, 0.1% lauryl sodium sulfate, 5 * SSC, the 20mmol/L sodium phosphate, the 0.5mg/mL salmon sperm DNA, 20mL,
2# liquid, hybridization solution contains 50% deionized formamide, 5 * Denhardt ' s, 0.1% lauryl sodium sulfate, 5 * SSC, the 20mmol/L sodium phosphate, the 0.5mg/mL salmon sperm DNA, 20mL also contains the prehybridization solution 20mL of 0.005-0.030mg/mL dna probe,
3# liquid; Cleaning fluid I# contains 2 * SSC of 0.1% lauryl sodium sulfate, 40mL,
4# liquid; Cleaning fluid II# contains 0.1 * SSC of 0.1% lauryl sodium sulfate, 40mL,
5# liquid; Damping fluid I# contains 100mmol/L trishydroxymethylaminomethane hydrochloric acid, 150mmol/L sodium chloride, and 100mL,
6# liquid; Damping fluid II# contains 100mmol/L trishydroxymethylaminomethane hydrochloric acid, 150mmol/L sodium chloride, and 3% (w/v) bovine serum albumin(BSA) 40mL,
7# liquid; Antibody liquid contains anti digoxin antibody in conjunction with protein 75 0U/mL, 0.005mL,
8# liquid; Damping fluid III# contains 100mmol/L trishydroxymethylaminomethane hydrochloric acid, 100mmol/L sodium chloride, and the 50mmol/L magnesium chloride, 40mL,
9# liquid; Contain chlorination nitroblue tetrazolium 32.5mg/mL, 5-bromo-4-chloro-3-indolylphosphate 17.5mg/mL, 0.2mL,
10# liquid; Colour developing liquid virus-positive contrast (CK+), 0.010mL,
11# liquid; Virus negative control (CK-), 0.010mL,
Hybond membrane (4cm * 10cm) 1,10 of hybrid pipes (5mL).
The hybridization detection kit of three kinds of above-mentioned viruses provides 10 times detection limit.
Pear virus hybridization detection kit and detection method thereof and step are as follows:
A. point sample: in advance film is cut into little of 1cm * 4cm, on film, performs mark with pencil, so that distinguish, at most can sample of every 0.5cm point (totally 7 samples).The sample RNA that extracts and positive control, negative control are got the 0.001mL point respectively to film,
B. fixing: with 80 ℃, 60 minutes or 120 ℃, 30 minutes fixed samples,
C. prehybridization: the film that fixes is put into a 5mL hybrid pipe, add 2mL1# liquid, 42 ℃ of prehybridizations 20 minutes,
D. probe is handled: the sex change 10 minutes in boiling water bath of 2# liquid, move to 5 minutes rapidly on ice,
E. hybridization: add 2mL2# liquid in the hybrid pipe, 42 ℃ of jogs 1.5 hours,
F. wash film: wash film 2 times with the 2mL3# room temperature, each 5 minutes; Wash film 2 times with under 42 ℃ of the 2mL4# liquid again, each 5 minutes,
G. sealing: washed film 1 minute with 2mL5# liquid, discard, add an amount of 2mL6# liquid jog sealing 20 minutes
G. sealing: washed film 1 minute with 2mL5# liquid, discard, add an amount of 2mL6# liquid jog sealing 20 minutes.
H. binding antibody: abandon 6# liquid, add 2mL 6# liquid (wherein adding 0.0004mL 7# liquid) again, jog 30 minutes,
I. colour developing: film is moved in the new pipe,, each 10 minutes, soak with 2mL8# liquid again and poured out in 2 minutes with 3mL5# liquid washed twice; Again after adding 2mL8# liquid (wherein adding 0.02mL9# liquid), container is put into the box of a sealing, leaves standstill colour developing 1 hour in the dark,
J. cessation reaction: wait to develop the color when suitable, pour out colour developing liquid, add 2mL5# liquid and soak 5 minutes cessation reactions,
The result observes and record:
Virus manifests the bluish violet spot at the point sample position.
The used probe sequence of said method is as follows:
The pear ring mosais virus probe sequence is shown in the sequence in the sequence table (＜210〉1).
The pear ring mosais virus probe sequence is shown in the sequence in the sequence table (＜210 〉).
The apple stem grooving virus probe sequence is shown in the sequence in the sequence table (＜210〉3).
The amount of above-mentioned probe is: 0.005-0.03mg/mL DNA.
The hybridization detection method of pear virus, testing sample or be the pear tree tissue, or organize RNA for pear tree.
Pear virus should manifest the bluish violet spot in the point sample position.
The reagent that said method is used, wherein deionized formamide, sodium phosphate, salmon sperm DNA, trishydroxymethylaminomethane, sodium chloride, bovine serum albumin(BSA), magnesium chloride 2, chlorination nitroblue tetrazolium, 5-bromo-4-chloro-3-indolylphosphate are provided by Huamei Bio-Engrg Co..Anti digoxin antibody is a ROCH company product in conjunction with albumen, digoxin-11-deoxyuridine triphosphate.The primer sequence that the dna probe of pear ring mosais, apple stem ditch, three kinds of viruses of pear vein yellow virus provides according to Shihezi Univ and at Shihezi Univ's mark digoxin-11-deoxyuridine triphosphate.The positive control (CK+) and the negative control (CK-) of pear ring mosais, apple stem ditch, three kinds of viruses of pear vein yellow virus are provided by Shihezi Univ.
The used instrument and equipment of said method comprises hybrid heater, constant temperature oven, refrigerator, microsyringe etc.
The present invention utilizes cDNA hybridization detection kit to detect on the pear tree 3 kinds of cryptovirus pear ring mosais, apple stem ditch, pear vein yellow virus.With respect to phyto-indicator method and serological detection method, have advantages such as good stability, highly sensitive, high specificity, quick, easy operating.Can be applied to the investigation and the virus-free standardization production of pears viral species and distribution situation efficiently.
Description of drawings
Accompanying drawing 1 is a pear vein yellow virus sample detection of the present invention image as a result.
Accompanying drawing 3 is an apple stem grooving virus sample detection of the present invention image as a result.
Test sample in the 1-10 in the diagram, the negative contrast of CK-.
Embodiment
Embodiment 1:
The hybridization detection kit and the detection method thereof of the pear ring mosais virus on the pear tree specifically comprise as follows:
The pear ring mosais virus probe sequence is shown in the sequence in the sequence table (＜210〉1).
The amount of above-mentioned probe is; 0.005-0.03mg/mL DNA.
1# liquid; Prehybridization solution contains: 50% deionized formamide, 5 * Denhardt ' s, 0.1% lauryl sodium sulfate, 5 * SSC, 20mmol/L sodium phosphate, 0.5mg/mL salmon sperm DNA, 20mL.
2# liquid; Hybridization solution contains, 50% deionized formamide, and 5 * Denhardt ' s, 0.1% lauryl sodium sulfate, 5 * SSC, the 20mmol/L sodium phosphate, the 0.5mg/mL salmon sperm DNA, 20mL also contains the prehybridization solution 20mL of 0.005-0.030mg/mL dna probe,
3# liquid; Cleaning fluid I# contains 2 * SSC of 0.1% lauryl sodium sulfate, 40mL,
4# liquid; Cleaning fluid II# contains 0.1 * SSC of 0.1% lauryl sodium sulfate, 40mL
5# liquid; Damping fluid I# contains 100mmol/L trishydroxymethylaminomethane hydrochloric acid, 150mmol/L sodium chloride, and 100mL,
6# liquid; Damping fluid II# contains 100mmol/L trishydroxymethylaminomethane hydrochloric acid, 150mmol/L sodium chloride, and 3% (w/v) bovine serum albumin(BSA), 40mL,
7# liquid; Antibody liquid contains anti digoxin antibody in conjunction with protein 75 0U/mL, 0.005mL,
8# liquid; Contain damping fluid III#, 100mmol/L trishydroxymethylaminomethane hydrochloric acid, 100mmol/L sodium chloride, the 50mmol/L magnesium chloride, 40mL,
9# liquid; Contain chlorination nitroblue tetrazolium 32.5mg/mL, 5-bromo-4-chloro-3-indolylphosphate 17.5mg/mL, 0.2mL,
10# liquid; Colour developing liquid virus-positive contrast (CK+), 0.010mL,
11# liquid; Virus negative control (CK-), 0.010mL,
Hybond membrane (4cm * 10cm) 1,10 of hybrid pipes (5mL).
The hybridization detection method and the step of the pear ring mosais virus virus of pear tree are as follows:
A. point sample: in advance film is cut into little of 1cm * 4cm, on film, performs mark with pencil, so that distinguish, at most can sample of every 0.5cm point (totally 7 samples).The sample RNA that extracts and positive control, negative control are got the 0.001mL point respectively to film,
B. fixing: with 80 ℃, 60 minutes or 120 ℃, 30 minutes fixed samples.
C. prehybridization: the film that fixes is put into a 5mL hybrid pipe, add 2mL1# liquid, 420 ℃ of prehybridizations 20 minutes,
D. probe is handled: the sex change 10 minutes in boiling water bath of 2# liquid, move to 5 minutes rapidly on ice,
E. hybridization: add 2mL2# liquid in the hybrid pipe, 42 ℃ of jogs 1.5 hours,
F. wash film: wash film 2 times with the 2mL3# room temperature, each 5 minutes; Wash film 2 times with under 42 ℃ of the 2mL4# liquid again, each 5 minutes,
G. sealing: wash film 1 minute with 2mL5# liquid, discard, add an amount of 2mL6# liquid jog sealing 20 minutes,
H. binding antibody: abandon 6# liquid, add 2mL 6# liquid (wherein adding 0.0004mL 7# liquid) again, jog 30 minutes,
I. colour developing: film is moved in the new pipe, with 3mL5# liquid washed twice, each 10 minutes, soak with 2mL8# liquid again and poured out in 2 minutes: after adding 2mL8# liquid (wherein adding 0.020mL9# liquid) again, container is put into the box of a sealing, and it is 1 little to leave standstill colour developing in the dark
J. cessation reaction: wait to develop the color when suitable, pour out colour developing liquid, add 2mL5# liquid and soak 5 minutes cessation reactions.
(3) result observes and record:
Virus manifests the basket purple dot at the point sample position.
The pear ring mosais virus probe sequence is shown in the sequence in the sequence table (＜210〉1).
The result observes and record:
Virus manifests the bluish violet spot at the point sample position.
Embodiment 2:
The difference of present embodiment and embodiment 1 is to be used for pear tree apple stem grooving virus probe sequence shown in the sequence (＜210〉2) of sequence table.
The amount of above-mentioned probe is: 0.005-0.03mg/mL DNA.
The result observes and record:
Virus manifests the basket purple dot at the point sample position.
Embodiment 3:
The difference of present embodiment and embodiment 1 is to be used for the pear vein yellow virus probe sequence shown in the sequence (＜210〉3) of sequence table.
The amount of above-mentioned probe is: 0.005-0.03mg/mL DNA.
The result observes and record:
Virus manifests the bluish violet spot at the point sample position.
Sequence table
＜110〉Shihezi Univ
＜120〉pear virus hybridization detection kit and detection method thereof
<160>9
<210>1
<211>629
<212>DNA
＜213〉pear ring mosais virus (pear ring pattern mosaic virus)
<400>1
tggaacagac?actggatgcc?atcttcgcga?acatagcgat?acaagggacg?tcggagcaga 60
cggaattcct?ggatctagtg?gtggaggtga?aatcaatgga?ggaccagaag?gtaatcgggt?120
cctacaattt?gaaggaggtg?gtcaacatga?tcaaagcttt?caaaactacc?tcttcggatc?180
cgaacatcag?caacatgact?ttccgccagg?tgtgtgaggc?tttcgcaccg?gaggcaagaa?240
acgggttggt?caagctgaag?tataaagggg?ttttcactaa?cctctttacg?accatgccgg?300
aagtaggtag?caaatacccg?gagctgatgt?ttgatttcaa?taagggtctt?aacatgttta?360
tcatgaataa?ggcccagcaa?aaagtcataa?ctaatatgaa?ccggcgtctt?ttacaaactg?420
aatttgcaaa?aagtgagaat?gaggcaaagc?tctcatctgt?tacgactgat?ctttgcattt?480
agtttgttta?agaagtttgg?tttaataaat?aaaataaata?gatagtgtgt?tgtgtgttta?540
atatttgcgt?gaatatatgt?ttgcatttaa?tggacgaact?cttgaaccca?tgaaagagta?600
taaagagtca?tggtatttaa?ttggagtgt 629
<210>2
<211>256
<212>DNA
＜213〉apple stem grooving virus (apple stem grooving virus)
<400>2
taggcagaac?tctttgaacg?aatgtacgtt?cagaaagctt?tgtgagcctt?ttgctgactt 60
ggctcgcgaa?tttcttcatg?agaggtggtc?cagaggattg?gccaccaaca?tttataagaa?120
atggcccaaa?gcttttgaaa?aaagcccatg?ggtggcgttt?gactttgcca?ccggtctgaa?180
aatgaatcgt?ttaacacctg?atgaaaaaca?ggtgattgat?agaatgacaa?aaaggctttt?240
tcgtactgaa?ggacaa 256
<210>3
<211>346
<212>DNA
＜213〉pear vein yellow virus (pear vein yellows virus)
<4000>3
tggaacctca?tgctgcaaac?tcagagtcct?cccgccaact?gggttggaaa?ggaattcaaa 60
ttcgaaacca?ggtatgccgc?ctttgacttc?ttctttggag?ttgagagctc?tgcatcgctg?120
gaaccagctg?atggcctcat?tcgattgcca?acccaggctg?aacgagttgc?caacgcaacg?180
agcaaagaaa?tccaaatgta?ccggattcga?tccatggagg?gaactcaggc?cgttaatttt?240
ggtgaggtta?cggggggcaa?agtgggccct?aagccagtgc?tgtccattag?gaagtaattg?300
atcttaaatc?ccataaacca?acaagtattt?atgcttttta?gtaaag 346
<210>4
<211>26
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>4
acactccaat?taaataccat?gactct 26
<210>5
<211>20
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>5
tggaacagac?actggatgcc 20
<210>6
<211>25
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>6
ttgtccttca?gtacgaaaaa?gcctt 25
<210>7
<211>25
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>7
taggcagaac?tctttgaacg?aatgt 25
<210>8
<211>30
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>8
ctttactaaa?aagcataaat?acttgttggt 30
<210>9
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>9
tggaacctca?tgctgcaaac?tc 22
Claims (2)
1. pear virus hybridization detection kit is characterized in that comprising:
The pear ring mosais virus probe sequence is as the sequence in the sequence table＜210〉shown in 1,
The apple stem grooving virus probe sequence is as the sequence in the sequence table＜210〉shown in 2,
The pear vein yellow virus probe sequence is as the sequence in the sequence table＜210〉shown in 3,
The consumption of above-mentioned probe is; 0.005-0.03mg/mL DNA,
1# liquid, prehybridization solution: contain: 50% deionized formamide, 5 * Denhardt ' s, 0.1% lauryl sodium sulfate, 5 * SSC, the 20mmol/L sodium phosphate, the 0.5mg/mL salmon sperm DNA, 20mL,
2# liquid, hybridization solution; Contain, 50% deionized formamide, 5 * Denhardt ' s, 0.1% lauryl sodium sulfate, 5 * SSC, the 20mmol/L sodium phosphate, the 0.5mg/mL salmon sperm DNA, 20mL also contains the prehybridization solution 20mL of 0.005-0.030mg/mL dna probe,
3# liquid, cleaning fluid I#: contain 2 * SSC of 0.1% lauryl sodium sulfate, 40mL,
4# liquid, cleaning fluid II: contain 0.1 * SSC of 0.1% lauryl sodium sulfate, 40mL,
5# liquid, damping fluid I#; Contain the 100mmol/L Tri(Hydroxymethyl) Amino Methane Hydrochloride, 150mmol/L sodium chloride, 100mL,
6# liquid, damping fluid II#: contain the 100mmol/L Tri(Hydroxymethyl) Amino Methane Hydrochloride, 150mmol/L sodium chloride, 3% (w/v) bovine serum albumin(BSA), 40mL,
7# liquid, antibody liquid; Contain anti digoxin antibody in conjunction with protein 75 0U/mL, 0.005mL,
8# liquid contains damping fluid III#; The 100mmol/L Tri(Hydroxymethyl) Amino Methane Hydrochloride, 100mmol/L sodium chloride, the 50mmol/L magnesium chloride, 40mL,
9# liquid contains chlorination nitroblue tetrazolium 32.5mg/mL, 5-bromo-4-chloro-3-indolylphosphate 17.5mg/mL, 0.2mL,
10# liquid, colour developing liquid virus-positive contrast (CK+), 0.010mL,
11# liquid, viral negative control (CK-), 0.010mL,
Hybond membrane 4cm * 10cm1 opens, and hybrid pipe 5mL10 props up.
2. pear virus hybridization detection method is characterized in that hybridization detection method and step are as follows:
A. point sample: in advance film is cut into little of 1cm * 4cm, on film, performs mark with pencil, so that distinguish, at most can sample of every 0.5cm point, totally 7 samples are got the 0.001mL point respectively to film with the sample RNA of extraction and positive control, negative control,
B. fixing: with 80 ℃, 60 minutes or 120 ℃, 30 minutes fixed samples,
C. prehybridization: the film that fixes is put into a 5mL hybrid pipe, add 2mL1# liquid, 42 ℃ of prehybridizations 20 minutes,
D. probe is handled: the sex change 10 minutes in boiling water bath of 2# liquid, move to 5 minutes rapidly on ice,
E. hybridization: add 2mL2# liquid in the hybrid pipe, 42 ℃ of jogs 1.5 hours,
F. wash film: wash film 2 times with the 2mL3# room temperature, each 5 minutes; Wash film 2 times with under 42 ℃ of the 2mL4# liquid again, each 5 minutes,
G. sealing: wash film 1 minute with 2mL5# liquid, discard, add an amount of 2mL6# liquid jog sealing 20 minutes,
H. binding antibody: abandon 6# liquid, add 2mL 6# liquid (wherein adding 0.0004mL 7# liquid) again, jog 30 minutes,
I. colour developing: film is moved in the new pipe,, each 10 minutes, soak with 2mL8# liquid again and poured out in 2 minutes with 3mL5# liquid washed twice; Again add 2mL8# liquid, wherein add 0.020mL9# liquid after, container is put into the box of a sealing, leave standstill in the dark colour developing 1 hour,
J. cessation reaction: wait to develop the color when suitable, pour out colour developing liquid, add 2mL5# liquid and soak 5 minutes cessation reactions,
The result observes and record:
Virus manifests the bluish violet spot at the point sample position,
Used probe sequence is as follows:
The pear ring mosais virus probe sequence is as the sequence in the sequence table＜210〉shown in 1,
The apple stem grooving virus probe sequence is as the sequence in the sequence table＜210〉shown in 2,
The pear vein yellow virus probe sequence is as the sequence in the sequence table＜210〉shown in 3,
The amount of above-mentioned probe is: 0.005-0.03mg/mL DNA.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100653066A CN100405061C (en) | 2005-04-01 | 2005-04-01 | Pear virus hybridization detection kit and detection method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100653066A CN100405061C (en) | 2005-04-01 | 2005-04-01 | Pear virus hybridization detection kit and detection method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1841067A CN1841067A (en) | 2006-10-04 |
| CN100405061C true CN100405061C (en) | 2008-07-23 |
Family
ID=37030170
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100653066A Expired - Fee Related CN100405061C (en) | 2005-04-01 | 2005-04-01 | Pear virus hybridization detection kit and detection method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100405061C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104805202A (en) * | 2015-04-24 | 2015-07-29 | 赛业（苏州）生物科技有限公司 | High-efficiency DNA hybridization solution and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1162979A (en) * | 1994-11-10 | 1997-10-22 | 拜尔公司 | DNA sequence and its use |
| WO2003093437A2 (en) * | 2002-05-02 | 2003-11-13 | University Of Rochester | Production of papillomavirus vaccines in plants |
| US20030213169A1 (en) * | 2002-05-14 | 2003-11-20 | Her Majesty In Right Of Canada As Respresented By The Minister Of Agriculture | Method of protecting plants from thrips |
| JP2004137239A (en) * | 2002-10-21 | 2004-05-13 | Bio Oriented Technol Res Advancement Inst | Agent and method for controlling soil blight |
| CN1503841A (en) * | 2000-06-13 | 2004-06-09 | 科技基因成果有限公司 | genes for s-adenosyl 1-methionine: jasmonic acid carboxyl methyltransferase and a method for a development of pathogen-and stress-resistant plants using the genes |
| WO2004112468A1 (en) * | 2003-06-17 | 2004-12-29 | Advanced Technologies (Cambridge) Limited | Plant limit dextrinase inhibitor |
-
2005
- 2005-04-01 CN CNB2005100653066A patent/CN100405061C/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1162979A (en) * | 1994-11-10 | 1997-10-22 | 拜尔公司 | DNA sequence and its use |
| CN1503841A (en) * | 2000-06-13 | 2004-06-09 | 科技基因成果有限公司 | genes for s-adenosyl 1-methionine: jasmonic acid carboxyl methyltransferase and a method for a development of pathogen-and stress-resistant plants using the genes |
| WO2003093437A2 (en) * | 2002-05-02 | 2003-11-13 | University Of Rochester | Production of papillomavirus vaccines in plants |
| US20030213169A1 (en) * | 2002-05-14 | 2003-11-20 | Her Majesty In Right Of Canada As Respresented By The Minister Of Agriculture | Method of protecting plants from thrips |
| JP2004137239A (en) * | 2002-10-21 | 2004-05-13 | Bio Oriented Technol Res Advancement Inst | Agent and method for controlling soil blight |
| WO2004112468A1 (en) * | 2003-06-17 | 2004-12-29 | Advanced Technologies (Cambridge) Limited | Plant limit dextrinase inhibitor |
Non-Patent Citations (2)
| Title |
|---|
| 库尔勒香梨的苹果茎痘病毒cDNA探针检测技术研究. 牛建新等.中国农业科学，第37卷第1期. 2004 |
| 库尔勒香梨的苹果茎痘病毒cDNA探针检测技术研究. 牛建新等.中国农业科学，第37卷第1期. 2004 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1841067A (en) | 2006-10-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Abravanel et al. | Performance of anti-HEV assays for diagnosing acute hepatitis E in immunocompromised patients | |
| Erensoy | Diagnosis of hepatitis C virus (HCV) infection and laboratory monitoring of its therapy | |
| CN103323590B (en) | Quantitative detection device based on fibrous-membrane gathering and separation and detection method thereof | |
| CN105734171A (en) | Zika virus fluorescent PCR detecting kit | |
| Majid et al. | Current and future hepatitis C virus diagnostic testing: problems and advancements | |
| CN112359137A (en) | RT-LAMP amplification system for visual virus nucleic acid RNA detection and application | |
| CN105671210A (en) | LAMP technology-based primer, kit and detection method for detecting Plum pox virus | |
| CN104388592B (en) | A kind of Porcine epidemic diarrhea virus S gene RT LAMP detection kits and detection method | |
| CN100405061C (en) | Pear virus hybridization detection kit and detection method thereof | |
| CN104212914B (en) | The heavy quick super quick detection kit of fluorescent PCR of Ebola five and application thereof | |
| CN104911254A (en) | Folding primer multiple PCR malaria molecule diagnostic kit and detection method thereof | |
| CN101875980B (en) | Kit and method for detecting Macrobrachium rosenbergii Nodavirus | |
| CN102758025A (en) | Reagent kit for fast detection of drainage oil and method for fast detection of drainage oil | |
| CN113913555B (en) | Detection card and detection kit for brassica flaviviruses | |
| CN101368964A (en) | Chemical luminescence method immune analysis diagnostic reagent kit for detecting cytomegalovirus IgG antibody | |
| CN103305635B (en) | Method for molecularly detecting sorghum mosaic virus infecting sugarcane | |
| CN114107561B (en) | GICA-RT-LAMP kit and detection card for detecting alfalfa mosaic virus | |
| CN105063238B (en) | A kind of RPA primers and kit for detecting grape leaf roll associated virus 2 | |
| Echevarría et al. | Sensitivity of a modified version of the ARCHITECT® Anti-HCV test in detecting samples with immunoblot-confirmed, low-level antibody to hepatitis C virus | |
| CN105624311A (en) | Real-time fluorescent PCR detection method and kit of multiple target nucleic acid in single tube | |
| CN105543350B (en) | A kind of the LAMP detection primer group and detection method of gnathostoma siamense | |
| CN110468224A (en) | Plasmodium specifically folds primer PCR molecule diagnosis kit and application method | |
| CN105002266A (en) | Universal method for carrying out malaria detection by adopting recombinase mediated isothermal nucleic acid amplification technology | |
| CN113846095B (en) | Detection card, detection primer composition and GICA-RT-LAMP kit for plantain mosaic virus | |
| CN103468816B (en) | Loop-mediated isothermal amplification (LAMP) primer and kit for shrimp composition and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080723 Termination date: 20100401 |