CN105010235A - Method for cleaning pig farm to prevent diseases - Google Patents
Method for cleaning pig farm to prevent diseases Download PDFInfo
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- CN105010235A CN105010235A CN201510503742.0A CN201510503742A CN105010235A CN 105010235 A CN105010235 A CN 105010235A CN 201510503742 A CN201510503742 A CN 201510503742A CN 105010235 A CN105010235 A CN 105010235A
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
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- Life Sciences & Earth Sciences (AREA)
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- Animal Behavior & Ethology (AREA)
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- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
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Abstract
The invention discloses a method for cleaning a pig farm to prevent diseases. Pig groups are divided into antibody negative subgroups and antibody positive subgroups according to immunity antibody detection; strengthen immunization is conducted on the antibody negative subgroups, immunity antibodies are detected again after immunization, and antibody positive pigs are brought into the antibody positive subgroups; fluid of umbilical cords of piglets is collected as detection samples when sows in the antibody positive subgroups give birth to the piglets, hog cholera virus and porcine pseudorabies virus are synchronously detected through a PCR method, and the pigs negative for both hog cholera virus and porcine pseudorabies virus are kept; regular casual inspection and comprehensive detection are conducted on the kept pigs, the pigs positive for both hog cholera virus and porcine pseudorabies virus or positive for one of hog cholera virus and porcine pseudorabies virus are weeded out, and then hog cholera virus and porcine pseudorabies virus are synchronously cleaned away. By synchronously cleaning away hog cholera virus and porcine pseudorabies virus, the method is high in efficiency, small in stress and easy and convenient to operate.
Description
Technical field
The present invention relates to disease field of purification, refer to the method for a kind of pig farm purification epidemic disease especially.
Background technology
Swine fever is commonly called as " rinderpest ", be the one that the Pestivirus suis belonged to by flaviviridae Pestivirus suis causes acute, heating, contagious disease, there is hyperinfection and lethal.Porcine pseudorabies is one of primary disease of swinery multiple infectious disease.The pig infected pigs pseudoabies of large-scale pig farm or swine fever directly or indirectly cause delivery room grice diarrhoea, the high inferior rate of child care pig, in the problem such as large porcine respiratory disease, cause high mortality and high incidence, bring huge economic loss to piggery.
In current large-scale pig farm, take the method purified respectively for swine fever and porcine pseudorabies.Purification for swine fever adopts collection tonsil to carry out swine fever purification usually, is carried out the purification of porcine pseudorabies subsequently by gene-deleted vaccine and discriminating.Its specific embodiments is: first, carries out immunity inoculation to kind of swinery, gathers sample and carries out detections immune antiboidy, and collection tonsil, carries out hog cholera immune fluorescence or Nest RT-PCR or fluorescence quantitative PCR detection Pestivirus suis, detect positive sow, eliminate.Subsequently, for PRV purification, namely immunity is in conjunction with antidiastole, detect that namely the wild malicious positive eliminates, the antidiastole sensitivity of current PRV virus is relatively low, usually needs 2-3 to monitor and positive boar superseded, finally reaches the object of purification.
Because different virus has different preferendums and the method for purification.The method of existing pig farm purification epidemic disease synchronously can not carry out the purification work of porcine pseudorabies and swine fever, need carry out respectively.And, because boar body weight is large, no matter gather vena cava anterior blood or gather tonsil all relatively complicated, and stress excessively easily cause farrowing sow to miscarry, blood specimen collection simultaneously needs many people to assist, and needing special installation, gatherer process is relatively time-consuming, gathers tonsil and easily occurs cross pollution.
Summary of the invention
In view of this, the object of the invention is to propose the method for a kind of pig farm purification epidemic disease, synchronous purification swine fever and PRV, efficiency be high, stress be little, easy to operate.
Based on the method for above-mentioned purpose pig farm provided by the invention purification epidemic disease, detected by immune antiboidy and be divided into negative antibody to hive off swinery and antibody positive hives off; Described negative antibody hives off and carries out booster immunization inoculation, again detects immune antiboidy after inoculation, and antibody positive pig is included described antibody positive in and hived off; Gather described antibody positive hive off middle Farrowing time piglet umbilical cord in body fluid as detection sample, synchronously detect Pestivirus suis and PRV by PCR method, retain Pestivirus suis and PRV is negative pig; Regularly inspect by random samples and complete detection the pig retained, superseded Pestivirus suis and PRV are the positive or one of them is positive pig, to realize the synchronous purification of Pestivirus suis and PRV.
Preferably, gather described antibody positive hive off middle Farrowing time piglet umbilical cord in body fluid as detection sample, extract DNA and RNA, the RNA wherein extracted respectively and detect Pestivirus suis by Nest RT-PCR method; The DNA extracted detects PRV by PCR method.
Preferably, the DNA of described extraction, by the gD genetic fragment of PCR method amplification PRV and gE genetic fragment, identifies through agarose gel electrophoresis.
Preferably, the size of described gD genetic fragment is 217bp, and the size of described gE genetic fragment is 350bp.
Preferably, described Nest RT-PCR method detects Pestivirus suis genetic fragment, and wherein overcoat PCR obtains the genetic fragment of 806bp, and it is 512bp that inner sleeve PCR obtains genetic fragment size.
Optionally, body fluid during described detection sample Farrowing in piglet umbilical cord is 3-5mL.
Preferably, what described immune antiboidy detected is antibody against swine fever virus, PRV gB antibody, pig annulus 2 type antibody and blue ear antiviral antibody, four kinds of immune antiboidies detect hives off entirely for positive pig includes antibody positive in, there is the wherein pig of one or more immune antiboidy feminine genders and includes negative antibody in and hive off.
Preferably, described negative antibody hives off and carries out booster immunization inoculation, inoculates it and is detected as vaccine corresponding to negative epidemic disease antibody once, inoculate latter one month, again carry out described immune antiboidy detection, detect positive pig and include described antibody positive in and hive off, negative pig is eliminated.
Preferably, in described regular sampling observation and complete detection, 20-30% sampling Detection antibody against swine fever virus, PRV gB antibody, pig annulus 2 type antibody and blue ear antiviral antibody is quarterly pressed once.
Preferably, in described regular sampling observation and complete detection, every half a year whole piglet break umbilical cord time gather body fluid in umbilical cord, detect Pestivirus suis and PRV by PCR method, and eliminate detection two kinds of epidemic diseases be the positive or wherein a kind of be positive pig.
As can be seen from above, the method for pig farm provided by the invention purification epidemic disease synchronously can reach purification swine fever and porcine pseudorabies two kinds of important diseases, easy and simple to handle, quick, without cross pollution and stress be little.Decrease the loss because swine fever and porcine pseudorabies cause.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet of the method for embodiment of the present invention pig farm purification epidemic disease.
Embodiment
For making the object, technical solutions and advantages of the present invention clearly understand, below in conjunction with specific embodiment, and with reference to accompanying drawing, the present invention is described in more detail.
Fig. 1 is the schematic flow sheet of the method for embodiment of the present invention pig farm purification epidemic disease.As shown in Figure 1, in the present embodiment, the method for described pig farm purification epidemic disease is detected by immune antiboidy and is divided into negative antibody to hive off swinery and antibody positive hives off; Described negative antibody hives off and carries out booster immunization inoculation, again detects immune antiboidy after inoculation, and antibody positive pig is included described antibody positive in and hived off; Gather described antibody positive hive off middle Farrowing time piglet umbilical cord in body fluid as detection sample, synchronously detect Pestivirus suis and PRV by PCR method, retain Pestivirus suis and PRV is negative pig; Regularly inspect by random samples and complete detection the pig retained, superseded Pestivirus suis and PRV are the positive or one of them is positive pig, to realize the synchronous purification of Pestivirus suis and PRV.Visible, the method for the present embodiment pig farm purification epidemic disease detects the umbilical cord body fluid of pig farm Farrowing, synchronous purification Pestivirus suis and PRV by PCR method, stress little, efficiency is high, easy to operate.
Embodiment one: immune antiboidy detects and tentatively hives off
Carry out a clinical appearance inspection to the boar of the whole audience, eliminate the boar of more than old, weak and farrowing 8 tires, composition supposition is healthy plants swinery.
Gather the vena cava anterior blood separation of serum that above-mentioned supposition health plants swinery, by ELISA (enzyme-linked immunosorbent assay) method detect supposition healthy plant swinery Pestivirus suis, PRV, pig circular ring virus, blue otopathy the expression of immune antiboidy (antibody against swine fever virus of biological Co., Ltd research and development before the section of Wuhan, PRV gB antibody, pig annulus 2 type antibody and blue otopathy poison antibody ELISA kit), choosing four kinds of antibody, to be positive pig be the swinery of successful immunization, includes antibody positive in and hive off.
Criterion:
Pestivirus suis: S/P (sample OD
630nmvalue/positive control OD
630nmvalue) value>=0.17, be judged to the positive; S/P value < 0.17, is judged to feminine gender.
PRV gB antibody: S/N (sample OD
630nmvalue/negative control OD
630nmvalue) value>=0.7, be judged to feminine gender; S/N value≤0.6 is positive; If 0.6 < S/N value≤0.7, for suspicious, sample detects again, if come to the same thing, then and interval two weeks sample detecting again.
Porcine circovirus 2 type antibody: sample (S-N)/(P-N) value >=0.16, be then judged to the positive; If sample (S-N)/(P-N) value < 0.16, be then judged to feminine gender.Wherein, S is sample OD
630nmvalue; P is positive control OD
630nmvalue; N is negative control OD
630nmvalue.
Blue ear antiviral antibody: sample KQ value >=20, are judged to the positive; Sample KQ value < 20, is judged to feminine gender.Wherein, KQ=(sample OD
630nmvalue/positive control mean value) * 100.
According to testing result, supposition health is planted swinery and be divided into antibody positive to hive off hiving off with negative antibody, two groups of isolated rearings.Wherein, it is positive pig that antibody positive hives off for above-mentioned immune antiboidy detects complete, and negative antibody hives off for above-mentioned detection exists the pig of immune antiboidy feminine gender.
Embodiment two: negative antibody hives off booster immunization
According to a kind of antibody test result of embodiment, the pig booster immunization that negative antibody hives off is inoculated it and is detected as vaccine corresponding to negative epidemic disease antibody once, according to corresponding vaccine operation instruction dose operation.
Inoculate corresponding vaccine after 1 month, again carry out corresponding immune antiboidy detection, based on embodiment one criterion, negative pig eliminated, positive pig is included antibody positive in and is hived off.
Embodiment three: antibody positive hives off synchronous purification
In the present embodiment, the qualified pig of antibody of above-mentioned antibody test result, namely antibody positive hives off pig, when Farrowing, gather the body fluid that piglet is broken in umbilical cord, deliver to laboratory, the body fluid that during Farrowing by collection, piglet is broken in umbilical cord, extracts DNA and RNA respectively.Wherein, the RNA of extraction detects Pestivirus suis by Nest RT-PCR method; The DNA extracted detects PRV by PCR method.The DNA of described extraction, by the gD genetic fragment of PCR method amplification PRV and gE genetic fragment, identifies through agarose gel electrophoresis.
Detect positive (carrier) pig, eliminate immediately after sow wean, piglet isolated rearing, booster immunization, no longer reserves seed for planting, and big porker is sold.
Test in laboratory concrete steps are as follows:
1) sample treatment
The body fluid 3-5mL that during Farrowing, piglet is broken in umbilical cord is as sample.Preferably, the body fluid 4mL in collection umbilical cord is as sample, and the centrifugal 15min of 1500rpm/min, gets supernatant 200 μ L, extracts DNA and RNA respectively.
2) RNA is extracted
Body fluid total serum IgE is slightly carried, and positive-virus is as positive control.
1. get pre-treatment Supernatant samples 200 μ L, be added in the Eppendorf pipe of 1.5mL, then add 1000 μ LRNA Trizol vibration mixings, room temperature leaves standstill 10min;
2. 200 μ L chloroforms are added, vibration mixing 5s (can not be too strong, in order to avoid produce emulsion layer, also can put upside down mixing with hand), ice bath 10min, 12000rpm/min 4 DEG C of centrifugal 10min on vortex mixer;
3. get supernatant in another new Eppendorf pipe, add equal-volume isopropyl alcohol (4 DEG C of precoolings) mixing, room temperature or 4 DEG C of standing 15min;
4. 4 DEG C, the centrifugal 10min of 12000rpm/min (Eppendorf tube opening keep towards centrifugal basket direction of principal axis place), carefully removes supernatant, is inverted in and blotting paper is stained with dry liquids or blots residual liquid with rifle, add 1mL 75% ethanol, put upside down washing;
5. the centrifugal 5min of 7500rpm/min, puts Eppendorf ramp down, exhausts liquid, air drying about 20min with the young heart of rifle tube.
6. be dissolved in the water that 25 μ L pollute without RNA enzyme ,-20 DEG C of short-term preservations ,-70 DEG C preserve for a long time or carry out reverse transcription immediately.
In the present embodiment, the utensil that all RNA extract reagent, various sample loading gun head, Eppendorf pipe and other contact samples must be all pollute without RNA enzyme, should wear disposable glove and often change during operation.
The reversion cDNA system (20 μ L) of RNA
System prepares to be placed in PCR instrument reacts, 42 DEG C, the extension of 60min, 70 DEG C, the deactivation of 5min reverse transcriptase.Reaction terminates, and by sample packing, puts into-20 DEG C of preservations after mark is good.
3) extraction of DNA
The blood sample that reference Tian Gen company provides, cell, tissue gene group DNA extraction kit specification carry out, and set up positive and negative to contrast.The DNA extracted is deposited 4 DEG C for subsequent use, if long storage time place-20 DEG C of preservations.
4) RT-PCR and pcr amplification:
1. Pestivirus suis Nest RT-PCR detection method mainly adopts Nest RT-PCR swine fever detection method, specific as follows:
Primer:
First, adopt Pup-1 and Pdown-1 to be respectively upstream and downstream primer, using above-mentioned steps 2) extract the body fluid total serum IgE of acquisition as template, reverse transcription carries out overcoat PCR reaction after obtaining cDNA.
Reaction system:
Amplification condition: 95 DEG C of 3min → (95 DEG C of 45s, 55 DEG C of 1mim, 72 DEG C of 1mim) 35 circulations → 72 DEG C extend 5min.
Subsequently, adopt Pup-2 and Pdown-2 to be respectively upstream and downstream primer, the PCR primer obtained with overcoat PCR, for template, carries out inner sleeve PCR reaction.
Reaction system:
Amplification condition: 95 DEG C of 3min → (95 DEG C of 45s, 55 DEG C of 1mim, 72 DEG C of 1mim) 35 circulations → 72 DEG C extend 5min.
2. PRV PCR detection method
First, with reference to pseudo-mad dog detection method (GB/T 18641-2002) amplification PRV gD genetic fragment, clip size is 217bp.
Primer:
Upstream primer Up1:5'-CACggAggACgAgCTggggCT-3'
Downstream primer Up2:5'-gTCCACgCCCCgCTTgAAgCT-3'
Reaction system:
Reaction condition:
95 DEG C of 5min → (94 DEG C of 1min, 57 DEG C of 1min, 72 DEG C of 1min) 30 circulations → 72 DEG C extend 10min, last 4 DEG C of protections, terminate reaction.
Secondly, according to PRV Becker pnca gene group sequence (GenBank:JF797219) the gene order design antidiastole PRV gE gene announced, object fragment is 350bp.
Primer:
Upstream primer gEup:5'-ATG CGG CCC TTT CTG CTG CY-3'
Downstream primer gEdown:5'-CGA CAC GGC GTC GCA GCC GC-3'
Reaction system:
Reaction condition:
95 DEG C of denaturation 5min, 94 DEG C of 1min, 60 DEG C of 1min, 72 DEG C of extensions, 30 circulations, last 72 DEG C of 10min.
5) agarose gel electrophoresis, analyses and comparison result
Get the product 10 μ l of PCR and Nest RT-PCR amplification, electrophoresis detection on 0.8% Ago-Gel.
Positive control and negative control are all set up, and then represent positive amplifying object fragment.
1. PRV:
Detect simultaneously the gE fragment positive of the gD genetic fragment of 217bp and 350bp for wild virus infection superseded;
GE genetic fragment only detected, in order between safe period, quarantine, blood sample collection carries out antidiastole;
Only detect that gD genetic fragment is then indicated as vaccine strain, do not eliminate.
2. Pestivirus suis: overcoat PCR obtains the genetic fragment of 806bp, it is 512bp that inner sleeve PCR obtains genetic fragment size, be then judged as the positive.
Culling level: detect that PRV gE and swine fever are positive or one of them is for positive, sow wean is superseded, and piglet isolated rearing, no longer reserves seed for planting, and eliminates.
Embodiment four: regularly sampling observation and complete detection
Maintain the kind swinery after said method purification, quarterly press 20%-30% sampling, detect the immune antiboidies such as Pestivirus suis, PRV, blue otopathy and PCV-II 1 time by embodiment one method.Antibody test feminine gender strengthens the immunity inoculation of vaccine, and the positive swinery of antibody test adheres to periodic detection and feeding and management.
Preferably, quarterly by 20% sampling.
Every half a year whole piglet break umbilical cord time, by the method for embodiment three, gather body fluid in umbilical cord, and detect Pestivirus suis and PRV, eliminate positive pig.Culling level: detect that PRV and swine fever are positive or one of them is for positive, sow wean is superseded, and piglet isolated rearing, no longer reserves seed for planting, and eliminates.
Through 1-2, carry out the monitoring purification work such as the detection-superseded-immunity of swine fever and porcine pseudorabies, swine fever, PRV can be controlled completely, set up healthy and stable kind swinery.
Embodiment five:
When implementing purification techniques, also to carry out implementing of supporting measures, taking Synthetical prevention technology.
For doing the purification of swine fever well, following several supplementary measures should be carried out:
1) isolated rearing strictly will be carried out in pig farm, the feeding and management system of " all in all out ".Except doing the immunity of hog cholera vaccine, PRV disease vaccine well, also to carry out the immunity inoculation of aftosa, blue otopathy and parvovirus.
2) carry out " five fixed work ", namely regularly sterilize, regular expelling parasite, regularly kills mouse, regular desinsection, regularly medicinal health-care.Implement every bio-security comprehensively.
3) do immunoprophylaxis well by the immune programme for children of science, improve the non-specific immunity of pig.
4) full price of feeding safe feed, the feed of moldy metamorphism of forbidding to feed.
5) improve feeding environment, pay attention to animal welfare.
6) to the new boar introduced, strict quarantine be carried out, introduce porcine pseudorabies viral disease and antibody against swine fever virus is negative or wild malicious negative antibody pig.After introducing, isolated rearing 2 months, blood drawing sample inspection, antibody or wild virus infection antibody are that negative patient mixes group feeding again and supports with this other pig.The same with other swinery, every half a year once checks, will strictly isolate for the positive pig of the infection detected, after vaccination, does growing and fattening pigs and raises sale.
7) bio-safety depth is set up
In the present embodiment, pig farm planted agent is also provided with disinfection station, terminal, ozone generator, wheel barnyard, draws pig car, air filtering boar station, independently feed factory specially.
Wherein, disinfection station construction and be applied as diseases prevention carry out first step measure; Terminal is isolated relative between each pig farm, between external client with pig farm, forms bio-safety depth first safety curtain; Ozone generator is march into the arena guarding of mouth in pig farm, comparatively utterly destroys important source of infection; The process that wheel barnyard is sudden illness and control and prevention of disease provide the space that can shift; Pig car is drawn to cut off transmission of pathogen between several field specially; Air filtering boar station provides the seminal fluid of high-quality; Independently feed factory reduces the risk owing to using outside feed.
8) reasonable immune programme for children is formulated
According to pig farm antibody test situation, in conjunction with pig farm Growth Results, clinical health degree suitably adjusts pig farm immune programme for children, guarantees pig farm administration measure and swinery health.
Embodiment six:
The kind pig farm of a scale 2000 sows, through carrying out the method for pig farm of the present invention purification epidemic disease from April, 10, carry out according to aforesaid operations, through the purification of 1.5 years, in October, 11, Pathogen test total negative, reached the clean-up effect of expection.And early stage adopts conventional method usually to need more than 2-3, and purification process stress be large, the problem such as cause that farrowing sow is miscarried.
Concrete Growth Results change sees the following form:
Because Pestivirus suis and PRV vertical transmission can occur, and umbilical cord connects parent and the unique passage of fetus, the developing immune system imperfection of fetus in addition, therefore after fetal infection virus, virus can be bred in fetus body, stays the body fluid in the fetus body in umbilical cord when body fluid is Farrowing in umbilical cord.Therefore the method for purification epidemic disease in pig farm of the present invention, swine fever and PRV virus infections can be judged whether by detecting body fluid in umbilical cord.Only gather a kind of sample, two kinds of diseases can be purified simultaneously, improve efficiency.
Meanwhile, for pseudoabies purification, the ELISA antidiastole technology for detection vena cava anterior serum gE antibody horizontal that tradition adopts, easily there is the problems such as false positive in the method, adopts round pcr effectively to avoid.
Visible, the break body fluid of umbilical cord of the method choice piglet of pig farm provided by the invention purification epidemic disease does Pestivirus suis and PRV for detecting sample, use the Nest RT-PCR technology and PRV PCR vaccine antidiastole technology that detect swine fever, positive pig detected, piglet isolated rearing is no longer reserved seed for planting, namely sow wean is eliminated, and detects-immunity-superseded program realization purification by 1-2.Avoid the shortcoming that conventional method need distinguish sample detecting purification, gather the purification that same sample can carry out Pestivirus suis and PRV simultaneously, improve purification efficiency.Further, the operability of sample collection is good, easier, quick compared with conventional Christmas operation, without cross pollution and stress be little.Avoid tonsil and forced venous blood sample collection complicated, easily pollute, stress be large, the problem such as easily cause that farrowing sow is miscarried.
Those of ordinary skill in the field are to be understood that: the discussion of above any embodiment is only exemplary, and not intended to be implies that the scope of the present disclosure (comprising claim) is limited to these examples; Under thinking of the present invention, also can combine between technical characteristic in above embodiment or different embodiment, step can realize with random order, and there are other changes many of different aspect of the present invention as above, and they do not provide in details for the sake of simplicity.Therefore, within the spirit and principles in the present invention all, any omission made, amendment, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a method for pig farm purification epidemic disease, is characterized in that, is detected be divided into negative antibody to hive off swinery and antibody positive hives off by immune antiboidy; Described negative antibody hives off and carries out booster immunization inoculation, again detects immune antiboidy after inoculation, and antibody positive pig is included described antibody positive in and hived off; Gather described antibody positive hive off middle Farrowing time piglet umbilical cord in body fluid as detection sample, synchronously detect Pestivirus suis and PRV by PCR method, retain Pestivirus suis and PRV is negative pig; Regularly inspect by random samples and complete detection the pig retained, superseded Pestivirus suis and PRV are the positive or one of them is positive pig, to realize the synchronous purification of Pestivirus suis and PRV.
2. the method for purification epidemic disease in pig farm according to claim 1, it is characterized in that, gather described antibody positive hive off middle Farrowing time piglet umbilical cord in body fluid as detection sample, extract DNA and RNA, the RNA wherein extracted respectively and detect Pestivirus suis by Nest RT-PCR method; The DNA extracted detects PRV by PCR method.
3. the method for pig farm according to claim 2 purification epidemic disease, is characterized in that, the DNA of described extraction, by the gD genetic fragment of PCR method amplification PRV and gE genetic fragment, identifies through agarose gel electrophoresis.
4. the method for purification epidemic disease in pig farm according to claim 3, it is characterized in that, the size of described gD genetic fragment is 217bp, and the size of described gE genetic fragment is 350bp.
5. the method for purification epidemic disease in pig farm according to claim 2, it is characterized in that, described Nest RT-PCR method detects Pestivirus suis genetic fragment, and wherein overcoat PCR obtains the genetic fragment of 806bp, and it is 512bp that inner sleeve PCR obtains genetic fragment size.
6. the method for purification epidemic disease in pig farm according to claim 1, it is characterized in that, body fluid during described detection sample Farrowing in piglet umbilical cord is 3-5mL.
7. the method for purification epidemic disease in pig farm according to claim 1, it is characterized in that, what described immune antiboidy detected is antibody against swine fever virus, PRV gB antibody, pig annulus 2 type antibody and blue ear antiviral antibody, four kinds of immune antiboidies detect hives off entirely for positive pig includes antibody positive in, there is the wherein pig of one or more immune antiboidy feminine genders and includes negative antibody in and hive off.
8. the method for purification epidemic disease in pig farm according to claim 1, it is characterized in that, described negative antibody hives off and carries out booster immunization inoculation, inoculate it and be detected as vaccine corresponding to negative epidemic disease antibody once, inoculate latter one month, again carry out described immune antiboidy detection, detect positive pig and include described antibody positive in and hive off, negative pig is eliminated.
9. the method for purification epidemic disease in pig farm according to claim 1, it is characterized in that, in described regular sampling observation and complete detection, quarterly press 20-30% sampling Detection antibody against swine fever virus, PRV gB antibody, pig annulus 2 type antibody and blue ear antiviral antibody once.
10. the method for purification epidemic disease in pig farm according to claim 1, it is characterized in that, in described regular sampling observation and complete detection, every half a year whole piglet break umbilical cord time gather body fluid in umbilical cord, detect Pestivirus suis and PRV by PCR method, and eliminate detection two kinds of epidemic diseases be the positive or wherein a kind of be positive pig.
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