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1.
Pseudorabies virus (PRV) is endemic in some regions of Japan. We investigated the effects of PRV infection status on herd productivity. Serum samples were obtained from 48 swine herds in Japan. Within each herd, three serum samples were obtained from growing pigs at four different ages, as well as from sows in low and high parity groups. Sera were tested for antibodies against wild-type PRV via competitive ELISA. Herds were classified into PRV positive and negative groups based on serological results. Herds infected with PRV exhibited postweaning mortalities (6.84%) that were significantly (P=0.0018) higher than those in unaffected herds (4.73%). Because of the reduced productivity in PRV positive herds, the current PRV eradication program must be strengthened.  相似文献   

2.
In theory, pseudorabies virus (PRV) may be eliminated from any size of breeding herd by phased test and removal if replacement gilts are not infected with PRV, culling decisions are partially based on PRV status, and the cull rate is higher than the incidence rate of PRV. Annual cull rates are commonly at least 50%, but little information exists on the incidence of PRV within enzootically infected swine herds. The purpose of this study was to develop a method by which spread of PRV could be detected among breeding swine within enzootically infected herds and to determine the incidence of PRV infection in these herds. Data were collected from 17 herds that were quarantined for PRV and ranged in size from 120 to 1,100 sows. At each herd, within the first 5 days of introduction, a group of approximately 30 replacement gilts was identified, vaccinated with a glycoprotein X-deleted PRV vaccine, and blood sample was collected. The owner of 1 herd had a nonvaccinated breeding herd and elected to leave incoming gilts nonvaccinated. After vaccination, blood samples were collected every 1 to 2 months for an average of 13.6 months. Serum samples from vaccinated gilts were tested for antiglycoprotein X antibodies by a specific differential ELISA. Samples from nonvaccinated gilts were evaluated by serum neutralization test. Product-limit method was used to estimate the probability of not becoming infected with PRV. Spread was detected in 7 of 8 herds that had more than 400 sows and in 2 of 9 herds that had less than 400 sows.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

3.
The potential of a pseudorabies virus (PRV) nucleocapsid protein (NC)-based enzyme-linked immunosorbent assay (ELISA) as a screening assay for PRV infection in subunit-vaccinated and nonvaccinated pigs was studied. The NC-ELISA compared favorably to a commercial ELISA for detecting PRV infection in nonvaccinated pigs. Virus-specific antibody was first detected by the NC-ELISA between days 14 and 21 in 5 pigs challenged intranasally with 10(4) PFU of virus. Antibody continued to be detected in these pigs through day 42, when the experiment was terminated. The NC-ELISA also detected antibody in 23 of 24 pigs from PRV-infected herds. In contrast, the commercial ELISA detected antibody 1 week earlier than the NC-ELISA in experimentally infected pigs but failed to detect antibody in 3 naturally exposed pigs that were identified by the NC-ELISA. Infection in these animals was confirmed by radioimmunoprecipitation analysis. The potential usefulness of the NC-ELISA for detecting infection in vaccinated pigs was also evaluated. The nucleocapsid-specific antibody responses of 10 PRV envelope glycoprotein subunit-vaccinated pigs were monitored prior to and following nasal exposure to a low dose (10(2.3) PFU) of PRV. Sera were collected periodically for 113 days after infection. Nucleocapsid-specific antibody responses measured by the NC-ELISA remained below the positive threshold before challenge but increased dramatically following virus exposure. Maximum ELISA responses were obtained on day 32 postchallenge (p.c.). Mean ELISA responses decreased thereafter but remained well above the positive threshold on day 113 p.c. PRV nucleocapsid protein can be used effectively as antigen in the ELISA for detecting PRV infection in both nonvaccinated and subunit-vaccinated pigs.  相似文献   

4.
Pseudorabies (Aujeszky disease) virus (PRV) was eliminated from domestic swine in many countries using glycoprotein E (gE)-deleted vaccines and serum antibody gE ELISAs, but PRV continues to circulate in some regions and in most feral swine populations in the world. We created a dual-matrix (serum and oral fluid) indirect IgG gE ELISA (iELISA) and evaluated its performance using samples from 4 groups of 10 pigs each: negative control (NC), vaccination (MLV), PRV inoculation (PRV), and vaccination followed by challenge (MLV-PRV). All serum and oral fluid samples collected before PRV challenge and all NC samples throughout the study were negative for gE antibodies by commercial blocking ELISA (bELISA) and our iELISA. Nasal swab samples from 9 of 10 animals in the PRV group were gB quantitative PRC (qPCR) positive at 2 days post-inoculation (dpi). The oral fluid iELISA detected a significant S/P response in the PRV (p = 0.03) and MLV-PRV (p = 0.01) groups by 6 dpi. ROC analyses of serum bELISA (n = 428), serum iELISA (n = 426), and oral fluid iELISA (n = 247) showed no significant differences in performance (p > 0.05). Our data support the concept of PRV surveillance based on oral fluid samples tested by an indirect gE ELISA.  相似文献   

5.
The use of an ELISA that can differentiate between swine infected with pseudorabies virus (PRV) and swine vaccinated with a specific PRV vaccine was evaluated on an individual and herd basis, and a system for interpreting ELISA results on a herd basis was developed. In 17 herds, recently introduced replacement gilts, seronegative for PRV, were vaccinated with a thymidine kinase- and glycoprotein X (gpX)-deleted vaccine. After vaccination, blood samples were collected from these gilts approximately every 1 to 2 months for up to 19 months. Serum samples were analyzed for antibodies to gpX antigen, using a commercially available ELISA kit according to the manufacturer's protocol. Herd status was determined as positive, suspect, or negative, according to the serum sample:negative control (S:N) values of the samples collected from the herd. From the 17 herds, 130 evaluations were performed. On 49 (38%) of the 130 herd evaluations, 1 or more gilts had suspect test results. Additional testing was required in 19 (39%) of these 49 herd evaluations to determine the PRV infection status of the herd. Status of herds having gilts with suspect results and no positive results was usually negative after retesting. Herds having gilts with positive results were unlikely to have negative status after retesting.  相似文献   

6.
A blocking enzyme-linked immunosorbent assay (ELISA) test has been developed to distinguish pseudorabies virus (PRV)-infected pigs from those immunized with a glycoprotein g92(gIII) deletion mutant, PRV(dlg92dltk). The blocking ELISA utilizes 96-well microtiter test plates coated with a cloned PRV g92(gIII) antigen, a mouse monoclonal antibody against gIII antigen (moMCAgIII): horseradish peroxidase (HRPO) conjugate, and undiluted test sera. Analyses can be completed in less than 3 hours with results printed out by an automated plate reader. Analyses on over 300 pig sera from PRV-free farms, on sera from other species, and on control sera containing antibodies to microorganisms other than PRV showed that the ratio of the optical density at 405 nm for the test sample to the optical density at 405 nm for the negative control (S/N value) was greater than 0.7 for all sera. No false positives were identified. Likewise, the S/N values were greater than 0.7 for over 400 sera obtained from pigs vaccinated twice with more than 1,000 times the standard PRV (dlg92dltk) dose or 1-4 times with the standard dose (2 x 10(5) TCID50/pig). Following challenge exposure to virulent PRV, the S/N values of the vaccinates were 0.1, showing that g92(gIII) antibodies in the sera of experimentally challenged pigs strongly blocked the binding of the moMCAgIII:HRPO conjugate to the antigen-coated wells. Sera of 233 pigs from PRV-infected herds with virus neutralization (VN) titers of 1:4 or greater were tested. All except 2 of these sera had S/N values less than 0.7 and more than 175 had S/N values less than 0.1. Sixteen sera from fetal pigs with VN titers of 1:4 or greater had S/N values of 0.24 or less, but 2 sera with VN titers of 1:4 when tested 5 years prior to the PRV g92(gIII) blocking ELISA test gave false negative S/N values. Twenty-four of 29 pig sera from PRV-infected herds with VN titers less than 1:4 were positive for g92(gIII) antibodies, illustrating the sensitivity of the PRV g92(gIII) blocking ELISA test. Analyses on 7 sera with VN titers of 1:4-1:64 showed that titers obtained with the PRV g92(gIII) blocking ELISA test were from 2- to 16-fold greater than the VN titers. The accuracy and sensitivity of the PRV g92(gIII) blocking ELISA test was further demonstrated by analyses of 40 unknown sera supplied in the National Veterinary Services Laboratories 1988 PRV check test kit.  相似文献   

7.
It has been reported that pseudorabies virus (PRV) stops spreading within growing-finishing sections of a large percentage of infected farrow-to-finish herds. This study was designed to follow the PRV status of growing-finishing pigs in a sample of infected herds. Fifteen infected herds were selected, of which 11 had seropositive finishing pigs and 4 had seronegative finishing pigs. These herds were visited quarterly for one year, and a cross section of growing-finishing pigs was tested for the presence of anti-PRV antibodies. The 4 herds that initially were seronegative remained seronegative, whereas of the 11 herds initially seropositive, 4 remained seropositive, 4 became seronegative, and 3 became temporarily seronegative before becoming seropositive again. Three characteristics serologic profiles were observed: one indicating continued viral spread; one indicating no spread for at least the preceding 3 months; and one indicating that PRV spread had recently ceased in this section of the herd. Results of our study indicated that periodic monitoring of a cross section of the growing-finishing pigs for their PRV serologic status was valuable for determining whether PRV was actively spreading in this section of the herd.  相似文献   

8.
从集贸市场随机采集97头份猪肉样本、53头份羊肉样本、50头份牛肉样本,应用双抗体夹心ELISA和PCR方法,平行检测样本可能含有的伪狂犬病病毒。结果97头份猪肉样中,双抗体夹心ELISA方法检出伪狂犬病病毒19份(19.6%),PCR检出25份(25.6%);牛肉样本中ELISA阳性数为0,PCR阳性2份(4%);羊肉样本中两种方法均为阴性。这两种方法均为阳性和阴性的份数为12和170,总符合率为89.2%。结果说明猪伪狂犬病的控制对提高肉食品品质具有重要作用。  相似文献   

9.
The objective of this study is the detection of Mycobacterium avium subsp. paratuberculosis (MAP) by serum enzyme-linked immunosorbent assay (ELISA), fecal polymerase chain reaction (PCR), and fecal culture in Colombian dairy herds. Serum and fecal samples from asymptomatic cows (n = 307) of 14 dairy herds were tested for MAP by an unabsorbed ELISA test (ELISA-A). Serum and fecal samples from positive ELISA-A animals (n = 31) were further tested by an absorbed ELISA test (ELISA-B) and PCR. Fecal samples from animals of herds positive by ELISA-A and PCR (n = 105) were inoculated onto three different culture media. ELISA-A produced positive results in 10% of the serum samples and 71% of the herds. ELISA-B and PCR results were positive in two and six serum and fecal samples from positive ELISA-A animals, respectively. Fecal samples were negative for MAP on all culture media. The results of this study confirmed the presence of MAP in local dairy herds and the difficulties of MAP detection in asymptomatic animals by ELISA, PCR, and fecal culture.  相似文献   

10.
A field evaluation of an enzyme-linked immunosorbent assay (ELISA) for swine trichinosis was done with sera obtained from 5 herds experiencing ongoing transmission of Trichinella spiralis. Epizootiologic studies conducted on these herds offered an opportunity to evaluate the accuracy of an ELISA, using larval T spiralis excretory-secretory antigens. Sera from 162 infected pigs and 143 serum samples from noninfected pigs originating from the same farms were tested. The infection status of the pigs was determined by digestion of diaphragm or tongue muscle samples. Two criteria were established to classify the ELISA optical density (OD) readings: Criterion I stated that an OD greater than or equal to 5 times the mean OD of several normal swine sera pools was positive; criterion II stated that a OD greater than or equal to 4 times the normal sera values was positive. The results obtained did not reveal obvious serologic variations among infected herds located in the 4 states involved. Overall, the test detected 93% (criterion I) and 96% (criterion II) of infected pigs. The majority of false-negative sera was from hogs that had less than 5 larvae/g of muscle; 1 hog had 73.8 larvae/g of diaphragm muscle. The false-positive rates were 8% for criterion I and 9% for criterion II. The actual rate for these false-positive samples may have been overestimated, because generally, only small tissue samples (0.4 to 10 g) were digested; larger sample sizes might have altered the results. The relevance of this qualification is that these pigs originated from herds with prevalence rates greater than 50%. Other factors that may account for occasional false-positive sera or false-negative sera in the swine trichinosis ELISA are discussed.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
Seroprevalence of porcine respiratory coronavirus in selected Korean pigs   总被引：8，自引：0，他引：8  
A total of 446 serum samples from 88 herds in Korea were examined for antibody to porcine respiratory coronavirus (PRCV) using blocking enzyme-linked immunosorbent assay (ELISA). All serum samples were collected from 24- to 26-week-old finishing pigs between December 1998 and June 1999. By ELISA, 237 out of 446 sera tested (53.1%) and 54 out of 88 sampled herds (61.3%) were positive against PRCV. Of 446 sera from 88 herd tested, 185 (41.5%) serum samples from 22 (25%) herds were seronegative against PRCV and transmissible gastroenteritis virus infection. Our data suggested that seropositive herds for PRCV are distributed diffusely throughout South Korea.  相似文献   

12.
The objective of this field study was to evaluate the protocol of test and removal (T&R) for the elimination of porcine reproductive and respiratory syndrome virus (PRRSV) from 5 chronically infected breeding herds. The T&R protocol involved sampling the entire breeding herd in one day, testing sera by polymerase chain reaction and ELISA to detect previously exposed and/or infected animals, and subsequently removing them from the herd. Following completion of T&R, breeding herds were monitored for 12 consecutive months, using ELISA, for the presence of antibodies to PRRSV. In order to be classified as a PRRSV-negative herd, all samples collected over the 12-month monitoring period were required to be negative by ELISA (s/p ratio < 0.4). At the conclusion of the monitoring period, all 5 farms were PRRSV-negative, according to the defined testing criteria. Approximately 2.2% (74/3408) ELISA false positive samples were detected across all 5 farms during the monitoring period. The diagnostic cost required during the T&R protocol was approximately US $10.66 per animal tested. Limitations of the study were a lack of herds with large (> 2000 sows) breeding herd inventories, and herds with a history of PRRSV vaccination.  相似文献   

13.
The validity of radial immunodiffusion enzyme assay (RIDEA) as a diagnostic test for antibodies to pseudorabies virus (PRV) in porcine serum was determined. Serum samples from sows and offspring were tested for the presence of antibodies to PRV, using both the RIDEA and the PRV serum-neutralization (SN) test. Overall sensitivity and specificity of the RIDEA done on serums from the sows were 95.7% and 95.2%, respectively. This sensitivity compares with 97.3% sensitivity of the SN test of the same serums. In 658 swine serum samples from routine submissions to the University of Missouri-Columbia Veterinary Diagnostic Laboratory that were tested by the RIDEA, the calculated sensitivity and the specificity were 94.3% and 98.9%. The RIDEA and SN test were equally sensitive (99.0%) to detect antibodies resulting from infection with a field strain of virus. They had reduced sensitivity (RIDEA, 91.7%; SN test, 95.2%) in tests of serums from vaccinated sows. For the detection of passively transferred antibodies in young pigs, sensitivity of the RIDEA was 76.1%, and specificity was 100%. In all instances, RIDEA was 100% sensitive at SN titers of 1:16 or greater. In testing serum samples of swine after field virus infection, sensitivity and specificity of the RIDEA approximated those of the SN test. This reliability, together with its ease of performance, makes the RIDEA an ideal field test in programs to detect PRV-infected herds and in programs designed to free herds of PRV infection.  相似文献   

14.
为了解湖南省规模化猪场猪伪狂犬病免疫状况及猪伪狂犬病毒感染情况,2018年在湖南省14个市州208个规模猪场采集4288份猪血清,采用ELISA方法检测血清中的伪狂犬病毒抗体。结果显示,免疫猪PRV gB抗体个体阳性率为78.1%,PRV gE抗体个体阳性率为21.0%,场阳性率为38.9%;从不同养殖规模来看,存栏量在1000头以上的大型猪场PRV gE抗体阳性率最高,为22.9%;在季节分布上,夏季猪群的PRV gE抗体阳性率最高,为25.1%;在不同的年龄阶段中,以种母猪PRV gE抗体阳性率最高,为36.2%。表明猪伪狂犬病在湖南地区仍广泛流行,野毒感染情况较为普遍。  相似文献   

15.
An indirect enzyme-linked immunoassay for serological surveillance of infection of pigs with Actinobacillus pleuropneumoniae (Ap) serotype 5 was developed. The antigen used was prepared from Ap serotype 5b strain L20. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the antigen contained high molecular weight lipopolysaccharide (LPS) and presumably also capsular polysaccharide (CP). The Ap serotype 5 ELISA was tested using sera from pigs experimentally infected with the 12 different Ap serotypes of biotype 1 and with sera from herds naturally infected with Ap serotypes 5, 6, 7 and 12. Cross-reactions were shown in one pig from a herd naturally infected with Ap serotype 7 and in one pig from a herd naturally infected with Ap serotype 12. The herd sensitivities of the Ap5 ELISA and a complement fixation test (CFT) were both estimated to 1.0, on the basis of serum samples from six herds naturally infected with Ap serotype 5. The herd specificities of both tests were estimated to 0.98, based on serum samples from 123 pig herds (10 samples from each herd) from the Danish specific pathogen-free (SPF) programme for pig production.  相似文献   

16.
In this study, we investigated whether Cedivac-FMD, an emergency vaccine against foot-and-mouth disease (FMD), is suitable for use conjointly with a screening program intended to confirm freedom from disease in vaccinated herds based on evidence of virus replication in vaccinates. Different sets of sera were tested using the Ceditest FMDV-NS ELISA for the detection of antibodies against non-structural proteins (NSPs) of FMD virus. During a vaccine safety study, serum samples were collected from 10 calves, 10 lambs and 10 piglets following administration of a double dose and a repeat dose of high payload trivalent Cedivac-FMD vaccine. All serum samples collected both 2 weeks following the administration of a double dose as well as those collected 2 weeks after the single dose booster (given 2 weeks after the double dose) were negative in the Ceditest FMDV-NS ELISA. In a series of vaccine potency experiments, serum samples were collected from 70 vaccinated cattle prior to and following exposure to infectious, homologous FMD virus. When testing cattle sera collected 4 weeks after vaccination with a regular dose of monovalent >6 PD(50) vaccines, 1 of 70 animals tested positive in the NSP antibody ELISA. After infection with FMD virus, antibodies to NSP were detected in 59 of 70 vaccinated cattle and 27 of 28 non-vaccinated control animals within 7 days. Cedivac-FMD vaccines do not induce NSP antibodies in cattle, pigs or sheep following administration of a double dose or a repeat dose. FMD-exposed animals can be detected in a vaccinated group within 7-14 days. Because Cedivac-FMD does not induce NSP antibodies, the principle of 'marker vaccine' applies.  相似文献   

17.
The introduction of Aujeszky's disease virus into a herd of pigs usually results in a rapid spread of the virus and a high percentage of pigs become seropositive. However, herd monitoring for the virus occasionally reveals a single seropositive breeding pig, referred to as a single reactor. The seropositive status of single reactors may be due to previous vaccination against Aujeszky's disease, or to exposure to a field strain of the virus, or to a false positive reaction in the serological assay. During a monitoring programme in Minnesota, 30 pig herds with single serological reactors were detected. Twenty-seven of these single reactors from 19 herds were segregated from their herds immunosuppressed with dexamethasone. Aujeszky's disease virus was isolated from four of the 27 pigs. Three of the four herds subsequently had outbreaks of Aujeszky's disease, suggesting that some single reactors were infected with Aujeszky's disease virus and had the potential to spread the virus within and between herds.  相似文献   

18.
The aim of the research was to assess the prevalence of antibodies to Coxiella burnetii in dairy cattle herds in Poland and to compare the results of real-time PCR and ELISA tests performed on bulk tank milk (BTM) samples. In total, 2635 serum samples collected from 969 dairy cattle herds from all provinces were tested using ELISA. Additionally, BTM specimens from 101 herds were analysed by ELISA and real-time PCR targeting IS1111 element. Presence of anti-C. burnetii antibodies was confirmed in 25.39% of serum samples in 237 herds (24.46%) and the herd-level seroprevalence in Voivodeships varied from 2.5% to 61.4%. Moreover, 46 (45.5%) of analysed bulk tank milk samples gave postive result in ELISA and microbial DNA was detected in 40 (39.6%) of tested herds. The comparative analysis of ELISA and real-time PCR results obtained for BTM samples using the chi-square test showed statistically significant relationship between results of both methods.  相似文献   

19.
Six 5-week-old pigs were inoculated intranasally (IN) with 10(7.6) TCID50 of bovine herpesvirus-1 (BHV-1). Three of the pigs also were inoculated IV with a similar dose of BHV-1. Clinical responses were not observed in these 6 pigs before oronasal challenge exposure with 10(7.8) TCID50 of virulent pseudorabies virus (PRV) at postinoculation day 42. Two pigs inoculated IN with BHV-1 and challenge exposed with PRV remained healthy, whereas the remaining 4 pigs developed severe clinical signs of pseudorabies and were moribund at postinoculation day 50 (8 days after challenge exposure). Anti-BHV-1 antibodies were demonstrable by ELISA in all 6 pigs and by serum neutralization (SN) in 5 pigs before challenge exposure with PRV. Anti-PRV antibody was not detected by ELISA or SN before challenge exposure to PRV. After challenge exposure to PRV, pigs with humoral antibody to BHV-1 responded anamnestically, and anti-PRV antibody activity was demonstrable by ELISA and SN in the 2 surviving pigs.  相似文献   

20.
于2009年3～5月采集上海地区的200份牛血清和400份猪血清,采用ELISA和荧光PCR方法检测其TTV（Torque Teno virus）感染状况。经ELISA检测,牛血清样品中检出阳性数2份,阳性率1．0％;猪血清样品中共检出阳性数15份,阳性率3．75％,其中生产母猪和育肥肉猪的阳性率分别为5．5％和2．0％,而荧光PCR检测结果均为阴性。结果表明上海地区猪和牛TTV感染现状处于较低水平。  相似文献   

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