CN107523627A - A kind of LAMP kit for detecting Acute Hepatic pancreatic necrosis cause of disease - Google Patents
A kind of LAMP kit for detecting Acute Hepatic pancreatic necrosis cause of disease Download PDFInfo
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- CN107523627A CN107523627A CN201710861917.4A CN201710861917A CN107523627A CN 107523627 A CN107523627 A CN 107523627A CN 201710861917 A CN201710861917 A CN 201710861917A CN 107523627 A CN107523627 A CN 107523627A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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Abstract
The invention discloses the LAMP primer group of one group of detection Acute Hepatic pancreatic necrosis cause of disease, it is made up of outer primer, inner primer and ring primer, the sequence such as SEQ ID NO of outer primer:1~SEQ ID NO:Shown in 2, the sequence such as SEQ ID NO of inner primer:3~SEQ ID NO:Shown in 4, the sequence such as SEQ ID NO of ring primer:5~SEQ ID NO:Shown in 6.The invention also discloses a kind of LAMP kit for detecting Acute Hepatic pancreatic necrosis cause of disease, the kit includes the LAMP primer group of above-mentioned detection Acute Hepatic pancreatic necrosis cause of disease, in addition to archaeal dna polymerase, 2 × reaction buffer, fluorescent dye, sealing fluid, standard positive template and negative control.Kit provided by the invention can detect Acute Hepatic pancreatic necrosis cause of disease directly from complex sample, with high specificity, high sensitivity, experimental repeatability is good, simple operation is quick the advantages of, expensive instrument is not needed to complete to detect, the scene available for Acute Hepatic pancreatic necrosis cause of disease is quick, accurately detection.
Description
Technical field
The present invention relates to technical field of microbial detection, in particular it relates to a kind of detection Acute Hepatic pancreatic necrosis cause of disease
LAMP kit.
Background technology
Penaeus Vannmei Deaths syndrome (EarlyMortality Syndrome, EMS) is defined as urgency in Thailand
Property Hepatopancreatic necrosis it is sick (Acute HepatopancreaticNecrosis disease, AHPND).It was found out from 2009
Begin, Acute Hepatic pancreatic necrosis (AHPND) breaks out in the world.But often recognized in early stage such case by raiser
Do not drawn attention to be fry quality problem.The disease is typically after seedling is thrown 20 to 30 days, and disease spreads rapidly, and the death rate shows
Write and rise.After morbidity, prawn typically exhibits：The symptom such as lethargic sleep, apocleisis, slow-growing, color is pale, soft shell.By Acute Hepatic
The prawn death rate may be up to 100% caused by pancreatic necrosis, very harmful to shrimp farming, the first half of the year in 2011, Hainan, Fujian,
The death loss of Guangdong and some areas of Guangxi is up to 80%.International food and agricultural organization FAO statistical result showeds in 2011, Malaysia
West Asia Penaeus Vannmei and Penaeus monodon yield total output dropped to 40000 tons by 70000 tons in 2010, the region of the disease
The extent of injury has been over white spot syndrome.In Vietnam, in June, 2011, cultivating penaeus monodon industry by unprecedented loss, its
Middle Tra Vinh saves 6200 hectare of cultivation by crushing blow, dead prawn sum about 3.3 hundred million tails, direct economic loss
12000000000 Dongs（Total about 3,800,000 RMB）；The north hectare of cultivation of village province 20000 is disaster-stricken, the Dong of economic loss 1,500,000,000,000
（About 4.7 hundred million RMB）.
Research data shows that AHPND cause of disease is a kind of Acute Hepatic pancreatic necrosis cause of disease for carrying the plasmid PVA1 that causes a disease
Specific strain.The PVA1 plasmids carry two encoding toxin protein genes, are separately encoded PirA and PirB toxin proteins.PirA
Prawn Hepatopancreatic necrosis can be caused with PirB toxin proteins, so as to which AHPND diseases occur.At present for PVA1 plasmids or
The regular-PCR detection method of PirA and PirB toxin proteins, but such method is high for equipment and personnel requirement, cost
It can be in any more, it is difficult to promote on a large scale.
Ring mediated isothermal amplification (loop-mediated isothermal amplification, LAMP) technology is a kind of
Novel isothermal nucleic acid amplification method, improved in the technology of traditional PCR technique, devise 4 or 6 it is different special
Property primer be directed to target gene 6 or 8 sites, at a kind of strand-displacement activity archaeal dna polymerase (Bst DNA polymerase)
In the presence of, realize DNA in constant temperature（65 DEG C or so）Under rapid amplifying, 10 can be reached in 1h9~1010Target sequence is copied
Shellfish.This method is simple to operate, and detection is quick, high sensitivity, and high specificity, it is not necessary to which expensive instrument can be completed to detect,
Opened in qualitative and quantitative detection, animal embryo sex identification and the genetic chip of the diagnosing of disease when participating in the cintest, popular bacterium or virus
It is used widely in the fields such as hair.Have not yet to see and LAMP technology is applied to Acute Hepatic pancreatic necrosis cause of disease in the prior artAP2In the detection of gene.
The content of the invention
The present invention is in order to overcome the above-mentioned deficiency of prior art, there is provided one group of detection Acute Hepatic pancreatic necrosis cause of disease
LAMP primer.
It is a further object to provide a kind of LAMP kit for detecting Acute Hepatic pancreatic necrosis cause of disease, the examination
Agent box can be directly complicated from matrix（Refer in food safety detection, background matrix exists to Testing index to be disturbed, and is often referred to host's base
Because of interference of group nucleic acid to detection target）Sample in detect Acute Hepatic pancreatic necrosis cause of disease, have high specificity, sensitivity
High, the advantages of experimental repeatability is good, simple operation is quick.
To achieve these goals, the present invention is achieved by following scheme：
The LAMP primer group of one group of detection Acute Hepatic pancreatic necrosis cause of disease, is made up of outer primer, inner primer and ring primer, draws outside
The sequence of thing such as SEQ ID NO:1~SEQ ID NO:Shown in 2, the sequence such as SEQ ID NO of inner primer:3~SEQ ID NO:4
It is shown, the sequence such as SEQ ID NO of ring primer:5~SEQ ID NO:Shown in 6.
The LAMP primer group is to be directed to Acute Hepatic pancreatic necrosis cause of diseaseAP2The specific primer group of gene design, by
A pair of outer primer F3（SEQ ID NO:1）、B3（SEQ ID NO:2）, a pair of inner primer FIP（SEQ ID NO:3）、BIP（SEQ
ID NO:4）With a pair of ring primer LF（SEQ ID NO:5）、LB（SEQ ID NO:6）Composition.
A kind of LAMP kit for detecting Acute Hepatic pancreatic necrosis cause of disease, comprising detecting acute hepatopancrease as described above
The former LAMP primer group of necrotic disease.
The kit also includes archaeal dna polymerase, 2 × reaction buffer, fluorescent dye, sealing fluid, standard positive template
And negative control.
Preferably, the mol ratio of the outer primer of primer sets, inner primer and ring primer is 1~2 in the kit：4~8：2
~4.
It is highly preferred that the mol ratio of the outer primer of primer sets, inner primer and ring primer is 1 in the kit：4：2.
Preferably, 2 described × reaction buffer is made up of buffer buffer solutions, glycine betaine and dNTPs, buffer bufferings
The volume ratio of liquid, glycine betaine and dNTPs is 10：8：7.
Preferably, the archaeal dna polymerase is Bst archaeal dna polymerases, and fluorescent dye is 0.02mM SYTO-9, and sealing fluid is
Mineral oil.
Preferably, the standard positive template is the matter containing Acute Hepatic pancreatic necrosis Pathogen test target gene AP2
Grain DNA, the negative control are sterilizing ultra-pure water.
It is highly preferred that the reaction system of the kit into being grouped into：Primers F 3, B3, FIP, BIP, LF, LB are anti-
It is respectively 0.2 μM, 0.2 μM, 0.8 μM, 0.8 μM, 0.4 μM, 0.4 μM to answer the final concentration in system, 2 × reaction solution 12.5 μ L, DNA
Polymerase 8U, 0.02mM SYTO-9 0.5 μ L, the μ L of measuring samples 2, add sterilizing ultra-pure water to 25 μ L；
The reaction condition of reaction system is 63 DEG C of 45~60min of reaction, and in 80 DEG C of lasting 2min.
Preferably, the detection method of the kit comprises the following steps：
S1. the processing of measuring samples；
S2.DNA preparation；
S3. ring mediated isothermal amplification detection reaction；
S4. testing result judges.
The present invention judges testing result according to amplification curve.Amplification curve is in serpentine, and testing result is the positive, that is, is examined
Contain Acute Hepatic pancreatic necrosis cause of disease in test sample product；Occur without serpentine amplification curve, testing result is feminine gender, that is, detects sample
Product do not contain Acute Hepatic pancreatic necrosis cause of disease.
Application of the kit in prawn Acute Hepatic pancreatic necrosis Pathogen test.
Compared with prior art, the invention has the advantages that：
（1）Specificity is good：The present invention is for one section of specific fragment that the AP2 target genes of design of primers are PVA1 plasmids, pin
Six special primers are designed to the purpose fragment to expand, high specificity, to prawn common virus, such as infectious subcutaneous and make
Blood organ necrosis virus（IHHNV）, prawn white spot syndrome disease（WSSV）, MBV（MBV）, shrimp bacterial liver
Pancreatic necrosis（NHPB）, prawn baculovirus（BP）, vibrio parahaemolytious DNA（Without PirA and PirB genes）In the absence of expansion
Increase.
（2）High sensitivity：For the positive plasmid containing Acute Hepatic pancreatic necrosis Pathogen test target gene AP2, most
Low detection limits can reach 10fg/ μ L；To actual positive sample, minimum detection limit can reach 1.12 × 10-4ng/μL。
（3）Experimental repeatability is high：Carry out 20 repetitions to negative control to test, primer does not expand；To positive control（Matter
Grain concentration 10pg/ μ L and 1pg/ μ L）Carry out 30 repetitions respectively to test, the Ct value coefficient of variation≤5%；Actual positive sample is entered
12 repetitions of row are tested, and amplification curve is in typical " S " type curve.
（4）Simple operation：Expensive, accurate equipment need not be used, it is low to instrument requirements, it is only necessary to a steady temperature
Instrument just can react and detect, and condition is fairly simple.
（5）Identification is simple：Acute Hepatic pancreatic necrosis cause of disease can be detected directly from complex sample, it is bent by observing amplification
Line directly judges yin and yang attribute, it is not necessary to other any analytical procedures such as cumbersome electrophoresis, is adapted to scene quickly, accurately to detect.
Brief description of the drawings
Fig. 1 is the negative control repeatability result schematic diagram that LAMP methods detect AHPND cause of diseases in application examples 1.
Fig. 2 is the positive control sensitivity results schematic diagram that LAMP methods detect AHPND cause of diseases in application examples 2.
Fig. 3 is the positive control stability result schematic diagram that LAMP methods detect AHPND cause of diseases in application examples 3.
Fig. 4 is the actual sample result schematic diagram that LAMP methods detect AHPND cause of diseases in application examples 4.
Fig. 5 is the actual sample electrophoresis result schematic diagram that regular-PCR detects AHPND cause of diseases in application examples 4.
Fig. 6 is the actual sample specific outcome schematic diagram that LAMP methods detect AHPND cause of diseases in application examples 5.
Fig. 7 is the actual sample sensitivity results schematic diagram that LAMP methods detect AHPND cause of diseases in application examples 6.
Embodiment
The present invention is made with reference to Figure of description and specific embodiment and further being elaborated, the embodiment
It is served only for explaining the present invention, is not intended to limit the scope of the present invention.Test method used in following embodiments is such as without spy
Different explanation, is conventional method；Used material, reagent etc., unless otherwise specified, for the reagent commercially obtained
And material.
Embodiment 1
A kind of LAMP kit for detecting Acute Hepatic pancreatic necrosis cause of disease, constituent are：One group of acute hepatopancrease of detection is bad
The LAMP primer group of dead cause of disease, archaeal dna polymerase, 2 × reaction buffer, fluorescent dye, sealing fluid, standard positive template and the moon
Property control.
The LAMP primer group of the detection Acute Hepatic pancreatic necrosis cause of disease, is made up of outer primer, inner primer and ring primer,
Be the AP2 genes using Acute Hepatic pancreatic necrosis cause of disease plasmid PAV1 as target gene, using primer-design software Primer
Explorer 4.0 designs 6 LAMP primers including ring primer, by raw work bioengineering（Shanghai）Limited company
Synthesis, the sequence such as SEQ ID NO of outer primer:1~SEQ ID NO:Shown in 2, the sequence such as SEQ ID NO of inner primer:3~
SEQ ID NO:Shown in 4, the sequence such as SEQ ID NO of ring primer:5~SEQ ID NO:Shown in 6.
The mol ratio of the outer primer, inner primer and ring primer is 1：4：2.
2 described × reaction buffer is made up of buffer buffer solutions, glycine betaine and dNTPs, and each component volume ratio is 10：
8：7.
The archaeal dna polymerase is Bst archaeal dna polymerases, and fluorescent dye is 0.02mM SYTO-9, and sealing fluid is mineral oil.
The standard positive template is the DNA containing Acute Hepatic pancreatic necrosis Pathogen test target gene AP2, institute
Negative control is stated as sterilizing ultra-pure water.
The reaction system of the kit into being grouped into：Primers F 3, B3, FIP, BIP, LF, LB are in reaction system
Final concentration be respectively 0.2 μM, 0.2 μM, 0.8 μM, 0.8 μM, 0.4 μM, 0.4 μM, the μ L of 2 × reaction solution 12.5, archaeal dna polymerase
The μ L of 8U, 0.02mM SYTO-9 0.5, the μ L of measuring samples 2, add sterilizing ultra-pure water to 25 μ L；
The reaction condition of reaction system is 63 DEG C of 45~60min of reaction, and in 80 DEG C of lasting 2min.
The detection method of the kit comprises the following steps：
S1. the processing of measuring samples；
S2.DNA preparation；
S3. ring mediated isothermal amplification detection reaction；
S4. testing result judges.
Application examples 1
The negative control repeatability of kit detection AHPND cause of diseases described in embodiment 1：
1st, template to be checked prepares：Positive control and negative control.
2nd, ring mediated isothermal amplification detection reaction system and condition：
25 μ L reaction systems contain：The final concentration of primers F 3, B3, FIP, BIP, LF, LB in reaction system is respectively 0.2 μM,
0.2 μM, 0.8 μM, 0.8 μM, 0.4 μM, 0.4 μM, the μ of 2 × reaction solution 12.5 μ L, archaeal dna polymerase 8U, 0.02mM SYTO-9 0.5
L, the μ L of measuring samples 2, add sterilizing ultra-pure water to 25 μ L.It is 20 μ L that sealing fluid, which adds volume,.Wherein, negative control sets 20 times and put down
OK, positive control set 2 times it is parallel.
Centrifuged after the PCR pipe configured is mixed, 63 DEG C of 45~60min of reaction, and in 80 DEG C of lasting 2min.
3rd, testing result judges：Above-mentioned reaction tube is placed in fluorescent PCR instrument（BIO-RAD）In, sentenced according to amplification curve
Disconnected testing result.Amplification curve is in serpentine, and testing result is the positive, that is, is detected in sample containing Acute Hepatic pancreatic necrosis disease
It is former；Occur without serpentine amplification curve, testing result is feminine gender, that is, detects sample and do not contain Acute Hepatic pancreatic necrosis cause of disease.
As shown in Figure 1, the results showed that：It is negative in the LAMP kit of the detection Acute Hepatic pancreatic necrosis cause of disease of foundation
Control is repeated 20 times detection without expanding.
Application examples 2
The positive control sensitivity of kit detection AHPND cause of diseases described in embodiment 1：
By positive control plasmid DNA carry out 10 times of gradient dilutions, respectively with 1ng/ μ L, 100pg/ μ L, 10pg/ μ L, 1pg/ μ L,
Seven 100fg/ μ L, 10fg/ μ L, 1fg/ μ L gradient concentration DNA are as template and negative control（Sterilize ultra-pure water）According to above-mentioned
Reaction system and condition establish detection method, to determine the sensitivity of kit.
As shown in Figure 2, the results showed that：After positive plasmid DNA is carried out into 10 times of gradient dilutions, the detection Acute Hepatic pancreas of foundation
The former LAMP kit of gland necrotic disease can detect the positive plasmid DNA that concentration is 10fg/ μ L.
Application examples 3
The positive control stability of kit detection AHPND cause of diseases described in embodiment 1：
Using the positive plasmid that concentration is 10pg/ μ L and 1pg/ μ L as template DNA, each set 30 times it is parallel, while feminine gender is set
Control（Sterilize ultra-pure water）Detection method is established according to above-mentioned reaction system and condition, to determine the stability of kit.
As shown in Figure 3, the results showed that, it is positive in the LAMP kit of the detection Acute Hepatic pancreatic necrosis cause of disease of foundation
Two concentration of plasmid repeat 30 progress test experiences, and repeatability is good, the Ct value coefficient of variation≤5%, it was demonstrated that the stabilization of kit
Property is good.
Application examples 4
It is as follows using the actual sample of the kit detection AHPND cause of diseases described in embodiment 1, step：
1st, the processing of measuring samples：The inward Vietnam grass shrimp live body hepatic tissue 2g of different batches is taken to be put into the APW of 10mL 3%（Chlorine
Change sodium basic peptone water）In culture medium, 37 DEG C of 8~18 h of culture carry out increasing bacterium.
2nd, DNA preparation：DNA is extracted from enrichment liquid using water-boiling method.
3rd, the DNA of extraction is detected according to above-mentioned reaction system and reaction condition, while negative control is set.
As shown in Figure 4, the results showed that：The detection Acute Hepatic pancreatic necrosis cause of disease established to inward Vietnam prawn
LAMP kit is detected, and typical " S " type curve amplification is presented in sample, is positive findings, it was demonstrated that detected in sample acute
Hepatopancreatic necrosis disease cause of disease.
Simultaneously using the actual sample of regular-PCR detection AHPND cause of diseases：
Using prawn live body hepatopancrease enrichment liquid DNA as template, enter performing PCR using primer AP3 and expand.Wherein, AP3 primers include upper
Swim primers F（SEQ ID NO:7）With anti-sense primer R（SEQ ID NO:8）.
Reaction system is：10×PCR buffer（Mg2+Plus）2.5 μ L, AP3 upstream and downstream primers（Each 10 μm of ol/L）1μ
L, dNTP（10μmol/L）2 μ L, the μ L of Taq DNA polymerase 0.5, the nucleic acid-templated 5 μ L of extracting, water is added to mend to 25 μ L.
Expanded according to following procedure：94 DEG C of 5min, then carry out 32 circulations（94 DEG C of 30s, 53 DEG C of 30s, 72
℃ 40s）, 72 DEG C of 5min, 4 DEG C of insulations.PCR primer is subjected to gel electrophoresis, compared with Fig. 4 results, to determine to establish
Detection Acute Hepatic pancreatic necrosis cause of disease LAMP kit testing result uniformity.
As shown in Figure 5, the results showed that：Actual sample is detected using regular-PCR method, electrophoresis result display detection sample
There is band at 336bp in product, in the same size with land control stripes band, and it is the positive containing AHPND cause of diseases to determine sample,
Regular-PCR result is consistent with LAMP kit testing result.
Application examples 5
The actual sample specificity of kit detection AHPND cause of diseases described in embodiment 1：
With the method for application examples 4 respectively to containing infectious hypodermal and hematopoietic necrosis virus（IHHNV）, prawn white spot syndrome
Disease（WSSV）, MBV（MBV）, shrimp bacterial Hepatopancreatic necrosis disease（NHPB）, prawn baculovirus（BP）, it is secondary
Hemolysis vibrion DNA（Without PirA and PirB genes）And the sample DNA of AHPND cause of diseases is detected.
As shown in Figure 6, the results showed that, there is amplification curve in the sample of the only cause of disease containing AHPND, and remaining sample does not expand,
The LAMP kit specificity for the detection Acute Hepatic pancreatic necrosis cause of disease that display is established is good.
Application examples 6
The actual positive sensitivity of kit detection AHPND cause of diseases described in embodiment 1：
10 times of gradient dilutions are carried out to the actual positive sample DNA stostes of the cause of disease containing AHPND of concentration known, according to above-mentioned reaction
System and condition are detected, while set negative control, to determine detection sensitivity of the kit to actual sample.Such as figure
Shown in 7, the results showed that, using initial concentration as 1.12 × 102Ng/ μ L positive sample DNA carries out gradient dilution, the detection of foundation
The LAMP kit of Acute Hepatic pancreatic necrosis cause of disease can reach concentration to the detection sensitivity of actual sample as 1.12 × 10- 4ng/μL。
Claims (9)
1. the LAMP primer group of one group of detection Acute Hepatic pancreatic necrosis cause of disease, it is characterised in that by outer primer, inner primer and ring
Primer forms, the sequence such as SEQ ID NO of outer primer:1~SEQ ID NO:Shown in 2, the sequence such as SEQ ID NO of inner primer:3
~SEQ ID NO:Shown in 4, the sequence such as SEQ ID NO of ring primer:5~SEQ ID NO:Shown in 6.
2. a kind of LAMP kit for detecting Acute Hepatic pancreatic necrosis cause of disease, it is characterised in that the kit includes right
It is required that the LAMP primer group of the detection Acute Hepatic pancreatic necrosis cause of disease described in 1.
3. kit according to claim 2, it is characterised in that the kit also includes archaeal dna polymerase, 2 × reaction
Buffer solution, fluorescent dye, sealing fluid, standard positive template and negative control.
4. the kit according to Claims 2 or 3, it is characterised in that mole of the outer primer, inner primer and ring primer
Than for 1~2：4~8：2~4.
5. the kit according to Claims 2 or 3, it is characterised in that mole of the outer primer, inner primer and ring primer
Than for 1：4：2.
6. kit according to claim 3, it is characterised in that 2 described × reaction buffer by buffer buffer solutions,
Glycine betaine and dNTPs compositions, the volume ratio of three are followed successively by 10：8：7.
7. kit according to claim 3, it is characterised in that the archaeal dna polymerase is Bst archaeal dna polymerases, fluorescence
Dyestuff is 0.02mM SYTO-9, and sealing fluid is mineral oil.
8. kit according to claim 3, it is characterised in that the standard positive template is bad containing acute hepatopancrease
Dead sick Pathogen test target gene AP2 DNA, the negative control are sterilizing ultra-pure water.
9. kit according to claim 3, it is characterised in that the reaction system of the kit into being grouped into：
The final concentration of primers F 3, B3, FIP, BIP, LF, LB in reaction system is respectively 0.2 μM, 0.2 μM, 0.8 μM, 0.8 μM, 0.4 μ
M, 0.4 μM, 2 × reaction solution 12.5 μ L, archaeal dna polymerase 8U, 0.02mM SYTO-9 0.5 μ L, the μ L of measuring samples 2, add sterilizing super
Pure water is to 25 μ L；
The reaction condition of reaction system is 63 DEG C of 45~60min of reaction, and in 80 DEG C of lasting 2min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111850153A (en) * | 2020-08-28 | 2020-10-30 | 福建省水产研究所（福建水产病害防治中心） | Primer group for detecting prawn acute hepatopancreatic necrosis disease-vibrio parahaemolyticus and kit containing primer group |
| CN115948582A (en) * | 2022-11-02 | 2023-04-11 | 中国海洋大学三亚海洋研究院 | Method for detecting acute hepatopancreatic necrosis pathogen based on ERA technology and application |
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| CN105316422A (en) * | 2015-12-02 | 2016-02-10 | 中国科学院南海海洋研究所 | Kit for rapidly detecting pathogeny of acute hepatopancreatic necrosis disease of prawns and application of kit |
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- 2017-09-21 CN CN201710861917.4A patent/CN107523627B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105316422A (en) * | 2015-12-02 | 2016-02-10 | 中国科学院南海海洋研究所 | Kit for rapidly detecting pathogeny of acute hepatopancreatic necrosis disease of prawns and application of kit |
Non-Patent Citations (1)
| Title |
|---|
| JETNAPHANG KONGRUENG ET AL.: "LAMP assay to detect Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease in shrimp", 《AQUACULTURE INTERNATIONAL》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111850153A (en) * | 2020-08-28 | 2020-10-30 | 福建省水产研究所（福建水产病害防治中心） | Primer group for detecting prawn acute hepatopancreatic necrosis disease-vibrio parahaemolyticus and kit containing primer group |
| CN115948582A (en) * | 2022-11-02 | 2023-04-11 | 中国海洋大学三亚海洋研究院 | Method for detecting acute hepatopancreatic necrosis pathogen based on ERA technology and application |
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