CN107955839A - For detecting double PCR primer, detection method and the kit of 3 type of porcine circovirus 2 type and circovirus - Google Patents
For detecting double PCR primer, detection method and the kit of 3 type of porcine circovirus 2 type and circovirus Download PDFInfo
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- CN107955839A CN107955839A CN201711324169.2A CN201711324169A CN107955839A CN 107955839 A CN107955839 A CN 107955839A CN 201711324169 A CN201711324169 A CN 201711324169A CN 107955839 A CN107955839 A CN 107955839A
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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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
- C12Q1/701—Specific hybridization probes
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- 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/16—Primer sets for multiplex assays
Abstract
The present invention provides double PCR primer, detection method and the kit for detecting 3 type of porcine circovirus 2 type and circovirus.Kit provided by the invention designs double PCR primer according to the highly conserved specific sequence of detection 3 type of porcine circovirus 2 type and circovirus, establishes single tube, the detection method of synchronous detection 3 type of porcine circovirus 2 type and circovirus.Kit of the present invention has the characteristics that fast and convenient, high specificity, sensitiveness height, good reliability, large batch of sample analysis can be carried out at the same time, primary first-order equation just can antidiastole sample be porcine circovirus type 2 infection, or 3 type of pig circular ring virus infection, or both co-infection, monitoring, prevention and control for pig circular ring virus epidemic situation provide strong technical support, there is good application prospect.
Description
Technical field
The invention belongs to biological technical field, and in particular to one kind is used to detect 3 type of porcine circovirus 2 type and circovirus
Double PCR primer, detection method and kit.
Background technology
Pig circular ring virus (porcine circovirus, PCV) nothing for circovirus section Circovirus a kind of small
Cyst membrane, icosahedral symmetry, ring-type, covalence closed Single-stranded DNA virus.Virion diameter average out to 17nm, with rolling ring side
Formula is replicated, and is presently found minimum no cyst membrane DNA virus.Brain, Allan etc. think can be according to pig circular ring virus
Pathogenic, antigenic and nucleotide sequence pig circular ring virus can be divided into two kinds of genotype, i.e. 1 type of pig circular ring virus (PCV1)
With porcine circovirus 2 type (PCV2).PCV1 was used as a kind of pollutants identification in 1974 in PK cell cultures first, its is right
Pig is not pathogenic.PCV2 reported that it can cause the pig circular ring virus of pig related in the clinical setting in 1998 first
Disease (Porcine circovirus associated diseases, PCVAD), mainly causes the exhaustion of piglet multisystem to integrate
Levy (PMWS), pigskin inflammation nephrotic syndrome (PDNS), porcine respiratory syndrome (PRDC), sow breeding difficulty disease, Hypertrophic intestines
Inflammation, gangrenosum acne interstitial pneumonia, piglet congenital tremors etc., show as breathing, uropoiesis, enteron aisle, lymph, angiocarpy, nerve, numerous
The dysfunction of system and skin is grown, great economic loss is caused to global pig-breeding.Recently, it is a kind of new
3 type virus of pig circular ring virus be reported first in the U.S., full length viral genome 2.0kb, its gene code two is main
Albumen：Cap protein and Rep albumen, both are in opposite direction in nucleic acid chains, it can cause pigskin inflammation nephrotic syndrome, numerous
Grow the symptoms such as obstacle, heart and multisystem inflammation.
The common detection method of detection PCV mainly has virus purification, Serologic detection and PCR detections etc. at present.But disease
Poison separation, Serology test is time-consuming and laborious, sensitiveness is low and false positive easily occurs.And PCR detection method mainly with
Based on the single detection of a certain Virus Type, it is impossible to carry out a variety of Virus Types joint PCR detection, and detection sensitivity with
Specificity is also uneven.
Since PCV2 and PCV3 can trigger porcine circovirus associated diseases, if can establish a kind of fast and convenient, high specificity,
Sensitiveness is high, PCV2 the and PCV3 joint-detection discrimination methods of good reliability, and the cause of disease of accurate recognition pig infection is simultaneously suited the remedy to the case,
It is significant to pig-breeding industry.
The content of the invention
To solve problem above, the present invention is provided to detect the double PCR of 3 type of porcine circovirus 2 type and circovirus
Primer, detection method and kit, establish it is a kind of can at the same time antidiastole PCV2 and PCV3 virus double PCR detection side
Method, and with very high detection sensitivity and specificity, accurately antidiastole the cause of disease of pig infection can be gone out, so as to implement correct
Therapeutic scheme, retrieve economic losses.
First aspect present invention provides a kind of double PCR for being used to detect 3 type of porcine circovirus 2 type and circovirus and draws
Thing, the primer are dual according to the highly conserved specific sequence design of detection 3 type of porcine circovirus 2 type and circovirus
PCR primer, wherein, the detection primer of porcine circovirus 2 type, by SEQID NO：The forward primer of nucleotide sequence shown in 3 and
SEQ ID NO：The reverse primer composition of nucleotide sequence shown in 4, detects the detection primer of 3 type of pig circular ring virus, by SEQ
ID NO：The forward primer and SEQ ID NO of nucleotide sequence shown in 11：The reverse primer group of nucleotide sequence shown in 12
Into.
The double PCR primer of above-mentioned detection 3 type of porcine circovirus 2 type and circovirus is preparing detection pig circular ring virus
Product in application.Second aspect of the present invention provides a kind of for detecting the dual of 3 type of porcine circovirus 2 type and circovirus
PCR detection method, comprises the following steps：
1) genome DNA is prepared from measuring samples；
2) using the genomic DNA of preparation as template, using double PCR primer as claimed in claim 1, in same reaction
Double PCR amplification is carried out in system, obtains amplified production；
3) amplified production obtains testing result through agarose gel electrophoresis.
In an embodiment of the present invention, the double PCR for detecting 3 type of porcine circovirus 2 type and circovirus is anti-
It is 50 μ l to answer system cumulative volume, including：
1) 10 μM of SEQ ID NO：3、SEQ ID NO：4、SEQ ID NO：11、SEQID NO：Nucleotides sequence shown in 12
Arrange each 1 μ l；
2)10×ExPCR Bμffer 5.0μl；
3)dNTP Mixtμre 4.0μl；
4) 1.0 μ l of ExTaq archaeal dna polymerases；
5)RNase free ddH20 32.0μl；
6) the 4 μ l of DNA profiling of sample to be tested.
In an embodiment of the present invention, the double PCR for detecting 3 type of porcine circovirus 2 type and circovirus is anti-
Condition is answered to include：Enter circulation after 94 DEG C of pre-degeneration 4min；94 DEG C denaturation 30s, 56 DEG C annealing 30s, 72 DEG C extension 30s, 35
After circulation terminates；72 DEG C extend 10min eventually.
In an embodiment of the present invention, the double PCR for being used to detect 3 type of porcine circovirus 2 type and circovirus
Detection method, its detection sensitivity is up to 102Copy/μ L.
Third aspect present invention provides a kind of double PCR detection for being used to detect 3 type of porcine circovirus 2 type and circovirus
Kit, includes the double PCR primer of detection 3 type of porcine circovirus 2 type and circovirus as described in the first aspect of the invention,
Further include 10 × PCR buffer, dNTP, MgCl2, Taq enzyme, positive quality control product, negative quality-control product.
In an embodiment of the present invention, the double PCR for being used to detect 3 type of porcine circovirus 2 type and circovirus
Detection kit can specific detection identification 3 type of porcine circovirus 2 type and circovirus, it is impossible to detect transmissible gastroenteritis of swine
One or more in virus, Porcine epidemic diarrhea virus, pig bocavirus, PRRS virus, pig triangle coronavirus.
Fourth aspect present invention provides a kind of double PCR primer as described in the first aspect of the invention, such as present invention second
Dual PCR detection method described in aspect or the double PCR detection kit as described in third aspect present invention are in pig circular ring virus 2
Application in poison detection, wherein, the detection is not for the purpose of medical diagnosis on disease is treated.
Brief description of the drawings
Fig. 1 is the primer screening electrophoretogram of double PCR provided in an embodiment of the present invention detection, wherein, M DL2000DNA
Marks, 1-9 are respectively negative control, PCV2-2-F/R and PCV3-4-F/R, PCV2-1-F/R and PCV3-4-F/R, PCV2-2-
F/R and PCV3-3-F/R, PCV2-2-F/R and PCV3-5-F/R, PCV2-3-F/R and PCV3-4-F/R, PCV2-3-F/R with
PCV3-5-F/R, PCV2-2-F/R and PCV3-1-F/R, PCV2-2-F/R and PCV3-2-F/R, PCV2-2-F/R and PCV3-6-
F/R；
Fig. 2 is that the primer concentration of double PCR provided in an embodiment of the present invention detection optimizes electrophoretogram, wherein, M is
DL2000DNA marks, 1 is negative control, and 2-6 is the addition of PCV2 and 3 primers of PCV, and it is respectively 0.4 μ that it, which adds volume,
L、0.6μL、0.8μL、1.0μL、1.2μL；
Fig. 3 is the annealing temperature optimization electrophoretogram of double PCR detection provided in an embodiment of the present invention, wherein, M is
DL2000DNA marks, 1 is negative control, and 2-9 is different annealing temperature, be respectively 60.1 DEG C, 59.3 DEG C, 58.0 DEG C, 56.0
℃、53.5℃、51.6℃、50.3℃、49.6℃；
Fig. 4 is the electrophoretogram of the specific test of double PCR provided in an embodiment of the present invention detection, and wherein swimming lane 2 is pig
The electrophoresis result of circovurus type 2 infection, swimming lane 3 are the electrophoresis result of 3 type of pig circular ring virus infection, and swimming lane 4 is pig circular ring virus 2
The electrophoresis result of 3 type mixed infection of malicious 2 types and pig circular ring virus, 5,6,7,8,9 be respectively transmissible gastro-enteritis virus, pig stream
Row diarrhea virus, pig Sai Neijia paddy virus, PRRS virus, the electrophoresis result of pig fourth type coronavirus, display do not detect；
Fig. 5 is the electrophoretogram of the sensitivity tests of double PCR provided in an embodiment of the present invention detection, and wherein swimming lane 2-10 divides
Wei 109Copy/μ L, 108Copy/μ L, 107Copy/μ L, 106Copy/μ L, 105Copy/μ L, 104Copy/μ L, 103Copy/μ
L、102Copy/μ L, 101Copy/μ L.
Embodiment
As described below is the preferred embodiment of the present invention, it is noted that for those skilled in the art
For, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also considered as
Protection scope of the present invention.
Below in conjunction with attached drawing, the preferred embodiment of the present invention is described in detail.It is not specified in embodiment specific
The experimental method of condition, usually according to normal condition.In the embodiment of the present invention unless otherwise noted, agents useful for same and consumptive material are
Commercial goods.
Embodiment 1
Porcine circovirus 2 type and the screening of 3 type dual-PCR method specific primers and system optimization
1) porcine circovirus 2 type and 3 type double PCR design of primers
The porcine circovirus 2 type (Porcine circovirus 2) and 3 type of pig circular ring virus 2 announced according to GeneBank
(Porcine circovirus 3) two kinds of pathogenic bacteria gene sequences, its highly conserved specific sequence is obtained by sequence analysis
Row, and Primer5.0 Software for Design specific primer is utilized according to the specific and conserved sequence.Primer is in the farsighted Boxing in Beijing
Bioisystech Co., Ltd of section synthesizes, and particular sequence is shown in Table 1：
Table 1 is directed to the specific primer of 3 type of porcine circovirus 2 type and pig circular ring virus 2：
2) reaction system and reaction condition of porcine circovirus 2 type and 3 type double PCRs
The reaction system of porcine circovirus 2 type and 3 type double PCRs is shown in Table 2：
The reaction system of 2 porcine circovirus 2 type of table and 3 type double PCRs
| Reaction reagent | Add volume |
| 10 × PCR Buffer pcr amplification reaction liquid | 5.0μL |
| dNTP Mixture | 4.0μL |
| 10 μM of porcine circovirus 2 types and 3 type primers | Each 1.0 μ L |
| PCR amplification enzyme | 1.0μL |
| Nucleic acid-templated (porcine circovirus 2 type and 3 types) | Each 2.0 μ L |
| Moisturizing is extremely | 50μL |
Reaction condition：Enter circulation, 94 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C of extensions after 94 DEG C of pre-degeneration 4min
30s, 35 after circulation terminates, and 72 DEG C extend 10min eventually, after reaction according to conventional agarose gel electroresis appraisal.
As a result：As shown in Figure 1, PCV2-2-F/R and PCV3-3-F/R specific bands are brighter, clear, and therefore, screening
PCV2-2-F/R and PCV3-3-F/R identifies primer for porcine circovirus 2 type and 3 type double PCR of pig circular ring virus.
3) primer concentration of porcine circovirus 2 type and 3 type double PCRs optimizes
To the porcine circovirus 2 type and pig circular ring virus 3 type double PCR primer PCV2-2-F/R and PCV3- of screening gained
3-F/R does primer concentration Optimal Experimental, adds 10 μM of porcine circovirus 2 types toward above-mentioned PCR reaction systems respectively and 3 type primers are each
For 0.4 μ L, 0.6 μ L, 0.8 μ L, 1.0 μ L, 1.2 μ L, corresponding primer concentration is 0.08 μm of ol/L, 0.12 μm of ol/L, 0.16 μ
Mol/L, 0.2 μm of ol/L and 0.24 μm of ol/L, are expanded by above-mentioned PCR reaction conditions, after reaction according to conventional agar
Sugared gel electrophoresis identification.
The results are shown in Figure 2, when porcine circovirus 2 type and 3 type primer amount of pig circular ring virus are respectively 1.0 μ L, band
Brighter, clear, it is respectively 1.0 μ L to determine each primer addition of double PCR, i.e., primer concentration is that 0.2 μm of ol/L is optimal.
4) annealing temperature of porcine circovirus 2 type and 3 type double PCRs optimizes
The reference value provided according to Primer5.0 during design of primers, by annealing temperature gradient be set as 49.6 DEG C, 50.3
DEG C, 51.6 DEG C, 53.5 DEG C, 56.0 DEG C, 58.0 DEG C, 59.3 DEG C, 60.1 DEG C of this 8 gradients respectively at cDNA templates carry out PCR expansions
Increase.
As a result：As shown in figure 3, when annealing temperature is 56.0 DEG C, band is brighter, clear, and therefore, optimum annealing temperature is
56.0℃。
Embodiment 2
The performance verification of kit of the present invention
1) specific test
Using the multiple PCR method of foundation to transmissible gastro-enteritis virus (TGEV), Porcine epidemic diarrhea virus
(PEDV), pig Sai Neijia paddy viral (SVV), PRRS virus (PRRSV), pig fourth type coronavirus (PDCoV) are expanded,
And aqua sterilisa is selected to do negative control, the specificity of inspection institute's method for building up.
The results are shown in Figure 4, and the purpose fragment size of porcine circovirus 2 type is 511bp；The purpose of 3 type of pig circular ring virus
Clip size is 701bp；And porcine circovirus 2 type and 3 type mixed infection purpose fragment size of pig circular ring virus have two, respectively
For 511bp and 701bp；TGEV, PEDV, SVV, PRRSV, PDCoV virus are showed no specific band amplification.
2) sensitivity tests
511bp fragments and 3 type gene of pig circular ring virus that porcine circovirus 2 type amplifies are connected to pMD19-T carriers,
Escherichia coli DH5a is converted, picking positive bacterium colony culture, (Plasmid Mini Kit I, are purchased from using plasmid purification kit
OMEGA companies) prepare the plasmid of purifying and carry out nucleic acid quantification, copy number is calculated, according to 1010Copy/μ L~101Copy/μ
The concentration of L carries out 10 times of gradient dilutions, takes the Plasmid DNA of 2 each dilution factors of μ L to optimize as template by step 3) described respectively
Reaction system and reaction condition carry out double PCR amplification sensitivity technique.As a result understand that the dual-PCR method is minimum to examine
Measure the viral DNA of 100 copies.
The results are shown in Figure 5, using after 10 times of doubling dilutions of mixing plasmid of PCV2 and PCV3 as template, with what is established
Multiple PCR method is expanded, and amplification shows that PCV2 detection limits are 3.192 × 102Copy/minimum the inspections of μ L, PCV3
It is measured as 2.667 × 102Copy/μ L.
3) multiplex PCR repetitive test
Using the multiplex PCR of foundation, detection is repeated once week about with optimization postcondition, it is continuous to detect 3 times, with detection
As a result reliability.With the mixing plasmid 10 of PCV2 and PCV38~101Copy/μ L are template, utilize the multiplex PCR side established
Method once, continuously expanded 4 weeks every amplification in 1 week.The result shows that 4 amplifications are the same, show the multiple PCR method have compared with
High repeatability.
The above embodiments are merely illustrative of the technical solutions of the present invention rather than limiting the scope of the invention, although ginseng
The present invention is explained in detail according to preferred embodiment, it will be understood by those of ordinary skill in the art that, can be to the present invention's
Technical solution technical scheme is modified or replaced equivalently, without departing from the spirit and scope of technical solution of the present invention.
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Claims (9)
- A kind of 1. double PCR primer for being used to detect 3 type of porcine circovirus 2 type and circovirus, it is characterised in that the primer Double PCR primer is designed according to the highly conserved specific sequence of detection 3 type of porcine circovirus 2 type and circovirus, wherein, The detection primer of porcine circovirus 2 type, by SEQ ID NO：The forward primer and SEQ ID NO of nucleotide sequence shown in 3：4 The reverse primer composition of shown nucleotide sequence, detects the detection primer of 3 type of pig circular ring virus, by SEQ ID NO：Shown in 11 Nucleotide sequence forward primer and SEQ ID NO：The reverse primer composition of nucleotide sequence shown in 12.
- 2. the double PCR primer of detection 3 type of porcine circovirus 2 type and circovirus is preparing detection as claimed in claim 1 Application in the product of pig circular ring virus.
- 3. a kind of dual PCR detection method for being used to detect 3 type of porcine circovirus 2 type and circovirus, comprises the following steps：1) genome DNA is prepared from measuring samples；2) using the genomic DNA of preparation as template, using double PCR primer as claimed in claim 1, in same reaction system Middle progress double PCR amplification, obtains amplified production；3) amplified production obtains testing result through agarose gel electrophoresis.
- 4. the dual PCR detection method as claimed in claim 3 for being used to detect 3 type of porcine circovirus 2 type and circovirus, its It is characterized in that, the reaction system cumulative volume of double PCR is 50 μ l, including：1) 10 μM of SEQ ID NO：3、SEQ ID NO：4、SEQ ID NO：11、SEQ ID NO：Nucleotide sequence is each shown in 12 1μl；2)10×ExPCR Bμffer 5.0μl；3)dNTP Mixtμre 4.0μl；4) 1.0 μ l of ExTaq archaeal dna polymerases；5)RNase free ddH20 32.0μl；6) the 4 μ l of DNA profiling of sample to be tested.
- 5. the dual PCR detection method as claimed in claim 3 for being used to detect 3 type of porcine circovirus 2 type and circovirus, its It is characterized in that, the program of double PCR includes：Enter circulation after 94 DEG C of pre-degeneration 4min；94 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C of extension 30s, 35 after circulation terminates；72 DEG C extend 10min eventually.
- 6. the dual PCR detection method as claimed in claim 3 for being used to detect 3 type of porcine circovirus 2 type and circovirus, its It is characterized in that, the detection sensitivity of the dual PCR detection method is up to 102Copy/μ L.
- A kind of 7. double PCR detection kit for being used to detect 3 type of porcine circovirus 2 type and circovirus, it is characterised in that bag The double PCR primer of detection 3 type of porcine circovirus 2 type and circovirus as claimed in claim 1 is included, further includes 10 × PCR buffer、dNTP、MgCl2, Taq enzyme, positive quality control product, negative quality-control product.
- 8. the double PCR detection kit as claimed in claim 7 for being used to detect 3 type of porcine circovirus 2 type and circovirus, It is characterized in that, the detection kit can specific detection identification 3 type of porcine circovirus 2 type and circovirus, it is impossible to examine Measure transmissible gastro-enteritis virus, Porcine epidemic diarrhea virus, pig Sai Neijia paddy virus, PRRS virus, pig fourth type hat One or more in shape virus.
- 9. a kind of double PCR primer as claimed in claim 1, dual PCR detection method as claimed in claim 3, such as weigh Profit requires application of the double PCR detection kit in pig circular ring virus detection described in 7, wherein, the detection is not with disease For the purpose of diagnoses and treatment.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108456747A (en) * | 2018-05-14 | 2018-08-28 | 湖北省农业科学院畜牧兽医研究所 | A kind of multiple PCR detection kit differentiating pig circular ring virus |
| CN108796124A (en) * | 2018-05-22 | 2018-11-13 | 广西壮族自治区兽医研究所 | A kind of the multiple PCR detection primer group and kit of quick differentiation PCV1, PCV2 and PCV3 |
| CN108998577A (en) * | 2018-09-18 | 2018-12-14 | 上海市动物疫病预防控制中心 | A kind of kit for detecting porcine circovirus 2 type and 3 types, primer pair, probe and method |
| CN109055619A (en) * | 2018-10-12 | 2018-12-21 | 华南农业大学 | For detecting double PCR primer, method and the kit of 3 type of pig circular ring virus and non-typical swine fever virus |
| CN109517929A (en) * | 2018-12-21 | 2019-03-26 | 武汉科前生物股份有限公司 | Primer sets and kit for pig circular ring virus detection and 2 type partings |
| CN109797248A (en) * | 2019-03-07 | 2019-05-24 | 长江大学 | A kind of universal porcine circovirus 2 type nested PCR detection method |
| CN111154915A (en) * | 2020-01-06 | 2020-05-15 | 咸阳职业技术学院 | PCR differential diagnosis kit for porcine circovirus type 4 and type 3 and detection method thereof |
| CN113046472A (en) * | 2019-12-27 | 2021-06-29 | 北京市农林科学院 | Triple PCR primer and kit for detecting porcine circovirus |
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| CN108998577A (en) * | 2018-09-18 | 2018-12-14 | 上海市动物疫病预防控制中心 | A kind of kit for detecting porcine circovirus 2 type and 3 types, primer pair, probe and method |
| CN109055619A (en) * | 2018-10-12 | 2018-12-21 | 华南农业大学 | For detecting double PCR primer, method and the kit of 3 type of pig circular ring virus and non-typical swine fever virus |
| CN109517929A (en) * | 2018-12-21 | 2019-03-26 | 武汉科前生物股份有限公司 | Primer sets and kit for pig circular ring virus detection and 2 type partings |
| CN109797248A (en) * | 2019-03-07 | 2019-05-24 | 长江大学 | A kind of universal porcine circovirus 2 type nested PCR detection method |
| CN113046472A (en) * | 2019-12-27 | 2021-06-29 | 北京市农林科学院 | Triple PCR primer and kit for detecting porcine circovirus |
| CN113046472B (en) * | 2019-12-27 | 2023-03-28 | 北京市农林科学院 | Triple PCR primer and kit for detecting porcine circovirus |
| CN111154915A (en) * | 2020-01-06 | 2020-05-15 | 咸阳职业技术学院 | PCR differential diagnosis kit for porcine circovirus type 4 and type 3 and detection method thereof |
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