规模化猪场猪圆环病毒2型感染的实验室诊断

为了确诊贵州省某规模化猪场保育仔猪异常死亡原因,从怀孕母猪、产房母猪、后备母猪、种公猪、哺乳仔猪、保育仔猪6个猪群采集90份血清样本采用ELISA方法分别进行血清抗体检测,并对采集的90份血清样本和1份病死猪淋巴结组织采用荧光PCR方法进行病原学检测。结果:6个猪群综合的猪瘟病毒、猪蓝耳病病毒、伪狂犬病病毒g B蛋白、伪狂犬病病毒g E蛋白血清抗体阳性率分别为87.78%、70.00%、88.89%、4.44%;猪瘟病毒、猪蓝耳病病毒、伪狂犬病病毒病原核酸检测显示阴性,猪圆环病毒2型病原核酸检测淋巴结组织样本显示阳性。试验结果表明,引起该猪场保育仔猪死亡的原因为猪圆环病毒2型感染。同时,怀孕母猪群出现了伪狂犬病病毒g E蛋白抗体阳性,提示怀孕母猪群可能存在猪伪狂犬病野毒感染。     

广西猪伪狂犬病毒感染状况调查报告

为了解广西猪伪狂犬病毒的感染状况,应用PCR技术对来自广西46个种猪场的1940份公猪精液进行了猪伪狂犬病毒(PRV)的检测,阳性率为0;同时对广西25个已使用猪伪狂犬病病毒gE基因缺失疫苗猪场的1455份送检血清,采用了猪伪狂犬病基因缺失鉴别ELISA方法进行了血清学抗体监测,结果检出猪伪狂犬gE抗体阳性11份,阳性率为0.76%。结果表明,广西猪群中存在猪伪狂犬病野毒感染,但感染率较低。     

应用ELISA检测猪伪狂犬病病毒抗体

应用 ELISA 对广西八个地区25个养猪点的380头份猪血清进行了猪伪狂犬病血清学调查.结果,在六个地区检出阳性22头份,阳性率为5.3%。从而证明了广西一些地区有伪狂犬病存在.用血清中和试验(SN)与 ELISA 对抽检的24份血清作了比较。结果,SN 和 ELISA 的阳性数分别为14份(58.3%)和20份(83.3%),ELISA 的敏感性高于 SN,且快速、简便.     

猪群进行猪伪狂犬病净化后病毒gE抗体的检测与分析

为了解猪伪狂犬病在规模化猪场的净化效果,采用ELISA对规模化猪场血清检测样品中猪伪狂犬病病毒gE抗体的水平进行判定。结果显示:净化前,3个规模化猪场共检测34份血清样品,其中猪伪狂犬病病毒g E抗体阳性3份,抗体阳性率达8.8%。采取净化措施后,3个猪场共检测34份血清样品,未检出猪伪狂犬病病毒gE阳性抗体,净化措施取得明显的效果。     

崇阳县猪伪狂犬病毒感染调查

为了解湖北省崇阳县猪伪狂犬病毒的感染状况,应用PCR技术对来自崇阳县不同规模7个种猪场的126份公猪精液进行了猪伪狂犬病毒(PRV)的检测,阳性率为0;同时采取了这7个已使用猪伪狂犬病病毒g E基因缺失疫苗免疫血清420份,采用了猪伪狂犬病基因缺失鉴别ELISA方法进行了血清学抗体监测,结果检出猪伪狂犬g E抗体阳性10份,阳性率为2.37%。结果表明,崇阳县猪群中存在猪伪狂犬病野毒感染,但感染率较低,不同猪场感染差异大。     

闽北地区猪伪狂犬病抗体检测分析

为了解猪伪狂犬病在闽北地区的流行情况,整理并分析2015年度血清抗体检测数据。采用ELISA检测方法对闽北地区45个规模化猪场进行检测分析,通过检测样品血清中伪狂犬病gE抗体和gB抗体的水平进行结果判定。结果显示：送检的45个规模化猪场,伪狂犬病gE抗体阳性场达25个,猪场阳性率达55.6%;共检测1304份血清,其中伪狂犬病gE抗体阳性213份、可疑24份、阴性1067份,抗体阳性率达20.6%。从伪狂犬病gE抗体阴性猪场抽检猪伪狂犬gB抗体380份,gB抗体阳性258份,猪群伪狂犬病抗体保护率仅有67.9%。     

比较两种ELISA试剂盒在猪伪狂犬病血清抗体检测中的一致性

比较两种ELISA试剂盒检测结果的一致性,为流行病学调查和临床诊断筛选适合的商品化试剂盒。采用来自两个厂家的野毒抗体(gE)和免疫抗体(gB)ELISA检测试剂盒分别检测362份和392份猪血清,应用Kappa检验比较试验数据的一致性。结果显示:两种猪伪狂犬病抗体(g E)检测试剂盒联合检测出95份抗体阳性和256份阴性,符合率97.24%,结果一致性为极强(Kappa值0.932);两种猪伪狂犬病抗体(gB)检测试剂盒联合检测出351份抗体阳性和9份阴性,符合率91.84%,一致性为弱(Kappa值0.347)。以上结果说明,两种猪伪狂犬病抗体(gE)检测试剂盒都适用于PRV gE抗体检测,而两种猪伪狂犬病抗体(gB)检测试剂金差异较大,需慎重选择。     

酒泉市猪场主要病毒性疾病的检测与分析

为了解2023年酒泉市几种主要猪病毒病的流行情况,本研究采用荧光定量RT-PCR方法,对酒泉市630份猪血清、270份猪组织进行了病原学检测。结果显示：检出阳性血清107份,样本个体阳性率由高到低依次为猪繁殖与呼吸综合征病毒(7.78%)、猪伪狂犬病病毒(4.92%)、猪细小病毒(4.29%)、非洲猪瘟病毒(未检出)、猪瘟病毒(未检出)、口蹄疫病毒(未检出);样本混合感染率为3.80%,猪繁殖与呼吸综合征病毒混合猪伪狂犬病病毒感染率最高(1.90%)。本次病原学检测可为酒泉市猪场疫病综合防控提供参考。     

达州市猪伪狂犬病的流行病学调查

为掌握达州市各地猪伪狂犬病的野毒流行情况,采用g E-ELISA方法随机抽检来自8个县122个场点的540份猪血清样本,共检出5个阳性县,30份猪伪狂犬病病毒(Pseudorabies Virus,PRV)阳性血清,阳性率为5.56%;检出14个阳性场,场点阳性率为11.48%,其中散养户场点阳性率最高,为15.15%。结果显示,达州市内大部分地区以及不同抽样群均存在不同程度的猪伪狂犬病野毒感染,提示应该进一步加强猪伪狂犬病的防控工作。     

2018年河南省郑州市定点监测猪群伪狂犬病gE抗体检测

为了解河南省郑州市2018年规模猪场的伪狂犬病野毒感染情况,利用gE抗体ELSIA方法,对免疫过猪伪狂犬病gE基因缺失疫苗的36个不同养殖规模的定点监测猪场,进行gE抗体检测。2018年1—12月,共检测血清样品1 057份,检出阳性场27个,场群阳性率为75.0%(27/36);检出阳性样品309份,样品阳性率为29.2%(309/1 057)。结果表明,郑州市伪狂犬病野毒感染面较广,流行率较高,存在较高的疫情暴发风险。存栏500头以下规模猪场的gE抗体阳性率(69.8%)是存栏3 000头以上的规模场(19.1%)的3.65倍,表明养殖规模越小,伪狂犬病野毒感染风险越大。利用荧光PCR方法对gE抗体阳性样品进行检测,检出阳性32份,阳性检出率为35.6%(32/90),表明感染猪群的持续带毒、排毒现象较为严重。建议规模化养猪场加强gE基因缺失疫苗免疫和gE抗体检测,及时淘汰阳性猪,同时结合隔离、消毒等综合防控技术,逐步实现猪伪狂犬病的控制与净化。     

石河子地区动物性食品中单核细胞增生李斯特菌的检测

目的了解新疆石河子地区动物性食品中单核细胞增生李斯特氏菌(LM)污染状况。方法在石河子地区选取5个具有代表性动物性食品零售点,对最常食用的生鲜猪肉、牛肉、羊肉、鸡肉、冻鸡肉、冻虾和冻带鱼8类动物性食品进行随机采样,采用病原分离培养和PCR法对样品中的单核细胞增生李斯特氏菌进行检测。结果检测8类249份食品样品,细菌分离鉴定阳性样品12份,平均阳性率4.82%;PCR法检测阳性样品36份,平均阳性率为14.46%。结论石河子动物性食品中LM的污染比较普遍,尤以冻鸡肉和冻虾LM污染较重,新鲜猪肉、牛肉和羊肉LM污染程度较轻。     

Detection of pseudorabies virus DNA in individual single-reactor pigs found in certified pseudorabies-free herds

During monitoring of certified pseudorabies (PRV)-free herds to confirm their PRV -free status, occasional individual gE-seropositive pigs are detected. These single-reactor pigs remain gE-seropositive when further serum samples are collected and tested. For the eradication programme to proceed, it is important to determine whether these pigs are only false positives or are; in fact, infected with field PRV. The purpose of this study was to determine whether the polymerase chain reaction (PCR) could detect field PRVDNA in single-reactor pigs and so confirm positive reactions in the serologic monitoring programme. First, DNA samples of various tissues from 15 single-reactor pigs all from different herds were examined for field PRV by PCR. Additionally, serum samples from these pigs were analyzed in a gE-confirmation enzyme linked immunosorbent assay (gE-confirmation ELISA). PCR detected PRVDNA in five of the 15 pigs, and these results were confirmed by the gE-confirmation ELISA. The remaining 10 pigs that tested negative in the PCR also tested negative in the gE- confirmation ELISA. We conclude that PCR can be used to discriminate between true and false serological positive single-reactor pigs and, moreover, that the gE-confirmation ELISA confirms these PCR results.     

青海省恰卜恰地区动物性食品中沙门菌的检测

对青海省恰卜恰地区的动物性食品——羊肉(20份)、牛肉(20份)、猪肉(10份)、鸡肉(10份)、牛奶(30份)进行了沙门菌的检验,各样品先用蛋白胨水或灭菌生理盐水增菌,然后在选择性培养基BS、HE培养该菌,经微量生化试验初步鉴定。结果显示,从90份样品中分离出3株沙门菌,总检出率为3.3%。     

我国主要畜牧业肉类产量的时空分析——基于空间自相关和重心轨迹的方法

利用GeoDa软件对我国猪肉产量、牛肉产量和羊肉产量的分布进行分析,得出以下结论：我国的猪肉产量、牛肉产量和羊肉产量都呈现上升的趋势,但它们的变动速率在不同的时期是不同的;我国猪肉产量、牛肉产量和羊肉产量都呈现聚集状态,猪肉产量的聚集显著性在上升,而牛肉产量和羊肉产量的聚集显著性在下降,猪肉产量主要聚集在南方,而牛肉产量和羊肉产量主要聚集在北方;猪肉产量的重心轨迹在湖北省境内向东南方向移动,牛肉产量的重心轨迹在山西省境内向西移动,羊肉产量的重心轨迹在陕西省境内向北移动。     

Detection of Porcine Circovirus Type 2, Porcine Parvovirus and Porcine Pseudorabies Virus from Pigs with Postweaning Multisystemic Wasting Syndrome by Multiplex PCR

Multiplex PCR was established to detect porcine circovirus type 2 (PCV-2), porcine parvovirus (PPV) and porcine pseudorabies virus (PRV) and applied to samples from 137 piglets exhibiting clinical signs of postweaning multisystemic wasting syndrome (PMWS). PCV-2 DNA was detected from all samples. Moreover, 43 samples were positive for PPV but negative for PRV; 11 samples were positive for PRV but negative for PPV; and 35 samples were positive both for PPV and PRV. These results suggests that PCV-2 co-infection with PRV and PPV may play an important role in PMWS. Also, multiplex PCR is an appropriate candidate method for diagnosis of PCV-2, PRV and PPV simultaneously in field cases.     

西宁市售动物性食品中肠出血型大肠杆菌O157：H7的检测

对西宁市售的动物性食品羊肉(20份)、牛肉(20份)、猪肉(20份)、鸡肉(20份)、牛奶(20份)、卤肉(10份)进行了大肠杆菌O157:H7检验,各样品先用mEC肉汤增菌,然后在选择性培养基TC-SMAC、MUG-LST培养此菌,经微量生化实验初步鉴定。结果是从110份样品中分离出1株大肠杆菌O157:H7,总检出率为0.9%;说明在西宁市售动物性食品中检出了大肠杆菌O157:H7。     

Identification of Pseudorabies Virus Variant and Control of Pseudorabies

Due to variant strain,pseudorabies virus (PRV) has broken out again and spread in China since 2011.A swine farm in Guangdong province was found pseudorabies (PR) symptoms-like miscarriage after introduction.The study was carried out to identify and control the PR.Serum of sows with and without miscarriage were randomly collected and the PRV gB and gE were detected by ELISA method,and brain tissues of sick piglets were sampled and the PRV gH gene was tested by PCR.All the sows in the farm were emergently inoculated PRV variant strainin activated vaccine.Serum before and after immunization were collected and detected by ELISA and micro-serum neutralization test.ELISA results showed that gE antibody of all the breeding sows with miscarriage were positive,and that of sows without miscarriage showed weekly positive;The average gB ELISA S/P value of sows with miscarriage was as high as 4.0,while that of sows without miscarriage was over 3.0.PCR of 3 sick piglets were all positive and the sequence of gB gene was 100% identical to BJ-YT-2012,a wide variant stain in 2012. The result of detection of the sows serum at before and after immunization showed that the S/P value of gB rose up from 1.603 before immunization to 2.88 at four weeks after immunization,and the neutralizing antibody rose up from 1:24 to 1:213.This agreed with the results that the sows showed less probability of miscarriage since the first week after immunization and almost no miscarriage after two weeks after immunization.This study suggested that classical PRV vaccine was not effective in this case,while the vaccine made from the variant PRV strain was.     

伪狂犬病病毒变异株的鉴定及伪狂犬病的防控

2011年以来伪狂犬病病毒（PRV）变异株在中国大范围流行致伪狂犬病（PR）再次暴发。广东某猪场发生疑似PR引起母猪较大范围的流产,为此本试验展开对该病诊断和防控方法的研究。随机抽取流产和未流产母猪血清,应用ELISA检测PRV gE和gB抗体;同时采集发病仔猪脑组织PCR检测PRV gH片段。对全场母猪紧急接种PRV变异株灭活苗,分别应用ELISA和中和试验检测免疫前后的血清抗体。结果显示,已发生流产母猪血清PR gE抗体均为阳性,而未流产母猪血清抗体见弱阳性;流产母猪PRV gB抗体的S/P值高达4.0,未流产母猪也达3.3。PCR检测3头病仔的脑组织均为阳性,测序表明其gB基因与2012年流行毒株BJ-YT-2012序列相似性为100%。ELISA检测免疫灭活疫苗前母猪血清PRV gB抗体S/P值为1.603,免疫4周后升高到2.88;特别是中和抗体从1：24升高到1：213。这与免疫疫苗1周后母猪流产开始减少,2周后母猪少见流产的结果吻合。研究结果提示,PRV经典株疫苗产生的PRV gB抗体对变异株的保护作用不佳,而变异株疫苗的保护效果显著。     

河北省猪场伪狂犬病病毒感染情况调查及gE基因遗传变异分析

采用ELISA检测方法对2018年1月至2019年5月从河北省11个地市195个不同规模猪场采集的10 479份血清进行gE抗体的检测,采用PCR方法对流产胎儿进行猪伪狂犬病病毒(pseudorabies virus,PRV)的抗原检測,并将部分阳性样本进行gE全基因的扩增和遗传变异分析.血清学检测结果显示,所检测的河...     

Technical note: A quick and more sensitive method to identify pork in processed and unprocessed food by PCR amplification of a new specific DNA fragment

We developed and evaluated a PCR procedure to detect pork in heated and unheated meat, sausages, canned food, cured products, and patés using a faster, more specific, and more sensitive method than others previously described. Isolation of a new DNA-specific porcine repetitive element was performed by nonspecific PCR amplification. After analyzing this repetitive sequence, a pair of primers were synthesized. To confirm the effectiveness and specificity of this fragment, 55 pig blood DNA samples (from differents breeds) were tested and positive results were obtained. With 200 samples tested from other species, the specific pork amplification was not detected. Using this method, we can partially quantify degree of contamination, depending on the PCR amplification cycles, detecting up to 0.005% pork in beef and 1% pork in duck paté using 30 and 20 PCR amplification cycles, respectively. The amount of porcine DNA detected in cattle DNA was 1.25 and 250 pg when using 30 and 20 amplification cycles, respectively. Pork has been identified in both heated and unheated meat products, sausages, canned food, hamburgers, and patés. In conclusion, specific PCR amplification of a repetitive DNA element seems to be a powerful technique for the identification of pork in processed and unprocessed food, because of its simplicity, specificity, and sensitivity (with 30 amplification cycles we can detect 0.005% pork). Furthermore, it is a very fast method, because 1% pork contamination can be detected with 20 PCR cycles. The procedure is also much cheaper than other methods based on RFLP-PCR, immunodiffusion, or other techniques that need expensive equipment.