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Porcine Pseudorabies Antibody ELISA kit, 192 Control Disease (Serum)

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[#AE-200130-2] Porcine Pseudorabies Antibody ELISA kit, 192 Control Disease (Serum)


AE-200130-2 | Porcine Pseudorabies Antibody ELISA kit, 192 Control Disease (Serum), 2x96 kit
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Pubmed ID :28982018
Publication Date : //
:  Feral swine ( Sus scrofa) are a pathogen reservoir for pseudorabies virus (PrV). The virus can be fatal to wildlife and contributes to economic losses in the swine industry worldwide. National surveillance efforts in the US use serology to detect PrV-specific antibodies in feral swine populations, but PrV exposure is not a direct indicator of pathogen transmission among conspecifics or to non-suid wildlife species. We measured antibody production and the presence of PrV DNA in four tissue types from feral swine populations of Florida, US. We sampled blood, nasal, oral, and genital swabs from 551 individuals at 39 sites during 2014-16. Of the animals tested for antibody production, 224 of 436 (51%) feral swine were antibody positive while 38 of 549 feral swine (7%) tested for viral shedding were quantitative polymerase chain reaction (qPCR)-positive for PrV. The detection of PrV DNA across all the collected sample types (blood, nasal, oral, and genital [vaginal] swabs) suggested viral shedding via direct (oronasal or venereal), and potentially indirect (through carcass consumption), routes of transmission among infected and susceptible animals. Fourteen of 212 seronegative feral swine were qPCR-positive, indicating 7% false negatives in the serologic assay. Our findings suggest that serology may underestimate the actual infection risk posed by feral swine to other species and that feral swine populations in Florida are capable of shedding the virus through multiple routes.

Authors : Hernández Felipe A, Sayler Katherine A, Bounds Courtney, Milleson Michael P, Carr Amanda N, Wisely Samantha M,

(2) A sensitive and specific lateral flow assay for rapid detection of antibodies against glycoprotein B of Aujeszky's disease virus.[TOP]

Pubmed ID :28919034
Publication Date : //
A direct double antibody lateral flow assay (DDA-gB-LFA) for the detection of antibodies against the glycoprotein B (gB) of Aujeszky's disease virus (ADV) in swine sera was developed. A native ADV gB was used for the preparation of a conjugate with colloidal gold particles and the immobilization on the strip membrane. The gB purified from ADV virions by immunoaffinity chromatography retained its native epitope structure after adsorption on the nitrocellulose membrane and the surface of colloidal gold particles. The diagnostic specificity and sensitivity of the DDA-gB-LFA were evaluated using 236 field swine sera. The diagnostic specificity and sensitivity of the DDA-gB-LFA compared to a commercially available gB-based ELISA were 98.0% and 98.6%, respectively, when determined with the use of the reader-detection mode, and 98.0% and 93.5%, respectively, when determined using visual detection. The DDA-gB-LFA provides a rapid, sensitive, and specific determination of ADV gB-directed antibodies in sera and can be used for the detection of ADV-exposed swine.

Authors : Vrublevskaya Veronika V, Afanasyev Vladimir N, Grinevich Andrey A, Skarga Yuri Y, Gladyshev Pavel P, Ibragimova Sagila A, Krylsky Dmitry V, Dezhurov Sergey V, Morenkov Oleg S,

(3) Enhancing expression of the pseudorabies virus glycoprotein E in yeast and its application in an indirect sandwich ELISA.[TOP]

Pubmed ID :28688204
Publication Date : //
The purpose of this study was to produce a recombinant pseudorabies virus (PRV) glycoprotein E (gE) protein with the correct antigenicity for use as a low-cost diagnostic antigen.

Authors : Wu C-Y, Wu C-W, Liao C-M, Chien M-S, Huang C,

(4) Development and Validation of Monoclonal Antibody-Based Antigen Capture ELISA for Detection of Group A Porcine Rotavirus.[TOP]

Pubmed ID :28414586
Publication Date : //
Porcine rotavirus-A (PoRVA) is one of the common causes of mild to severe dehydrating diarrhea, leading to losses in weaning and postweaning piglets. A rapid, highly specific, and sensitive antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for detection of PoRVA, by using VP6 (a highly conserved and antigenic protein of group-A rotavirus)-directed rabbit polyclonal antibodies (capture antibody) and murine monoclonal antibodies (detector antibody). The detection limit of AC-ELISA was found to be equal to that of conventional reverse transcription-polymerase chain reaction (RT-PCR; about 10 TCID/mL). For validation of the in-house AC-ELISA, 295 porcine fecal/diarrhea samples, collected from different provinces of China, were evaluated and compared with conventional RT-PCR and TaqMan RT-quantitative PCR (qPCR). The sensitivity and specificity of this in-house AC-ELISA relative to RT-qPCR were found to be 91.67% and 100%, respectively, with the strong agreement (kappa = 0.972) between these two techniques. Total detection rate with AC-ELISA, conventional RT-PCR, and RT-qPCR were found to be 11.2%, 11.5%, and 12.2%, respectively, without any statistical significant difference. Moreover, AC-ELISA failed to detect any cross-reactivity with porcine epidemic diarrhea virus, transmissible gastroenteritis virus, pseudorabies virus, and porcine circovirus-2. These results suggested that our developed method was rapid, highly specific, and sensitive, which may help in large-scale surveillance, timely detection, and preventive control of rotavirus infection in porcine farms.

Authors : Memon Atta Muhammad, Bhuyan Anjuman Ara, Chen Fangzhou, Guo Xiaozhen, Menghwar Harish, Zhu Yinxing, Ku Xugang, Chen Shuhua, Li Zhonghua, He Qigai,

(5) Development of a competitive double antibody lateral flow assay for the detection of antibodies specific to glycoprotein B of Aujeszky's disease virus in swine sera.[TOP]

Pubmed ID :27894861
Publication Date : //
Three lateral flow assays (LFAs) for the detection of antibodies against glycoprotein B (gB) of Aujeszky's disease virus (ADV) in swine sera: a competitive double antibody sandwich LFA without a preincubation step (CDAS-gB-LFA), a CDAS-gB-LFA with a preincubation step (pCDAS-gB-LFA), and a competitive direct gB-LFA have been developed and were compared with each other and with a gB-ELISA. The assays are based on monoclonal antibodies to immunodominant epitopes of ADV gB. The pCDAS-gB-LFA proved to be the most specific and sensitive assay to detect antibodies directed to ADV gB. The specificity and sensitivity of the pCDAS-gB-LFA with the use of an LFA reader for test line intensity measurements were 97.6 and 94.9%, respectively. The lower diagnostic sensitivity of the pCDAS-gB-LFA compared to a gB-ELISA reflects its reduced analytical sensitivity, which was shown in titration experiments with positive sera. The pCDAS-gB-LFA, using the reader-based and visual detection modes, showed good agreement in respect to specificity; however, the LFA reader detection provided a higher diagnostic and analytical sensitivity compared to visual detection. The developed pCDAS-gB-LFA is a rapid, sensitive, and specific method for the detection of antibodies to ADV gB and can be used for screening ADV-infected swine in unvaccinated herds.

Authors : Vrublevskaya V V, Afanasyev V N, Grinevich A A, Skarga Yu Y, Gladyshev P P, Ibragimova S A, Krylsky D V, Morenkov O S,

(6) Lactobacillus pentosus expressing porcine lactoferrin elevates antibacterial activity and improves the efficacy of vaccination against Aujeszky's disease.[TOP]

Pubmed ID :27653426
Publication Date : //
In this study, Lactobacillus pentosus expressing porcine lactoferrin (pLF) was tested for in vitro antibacterial activity and for its ability to enhance immunity induced by an orally administered Aujeszky's disease virus (ADV) vaccine. The cDNA encoding N-terminus of pLF was cloned into a Lactobacillus-specific plasmid to produce L. pentosus pLF expressing transformants (pPG612.1-pLFN/ L. pentosus). The antimicrobial activity of the recombinant pLF protein inhibited bacterial growth in vitro. The supernatant of pPG612.1-pLF-N/L. pentosus had an inhibitory effect on Staphylococcus aureus strain CVCC26003, Bacillus subtilis strain CVCC63501, Escherichia coli strain CVCC10141 and Salmonella enterica ssp. enterica Choleraesuis strain CVCC79102, while it did not inhibit the growth of Lactobacillus casei strain ATCC393. A mouse model was established to test the effectiveness of the orally administered probiotic L. pentosus recombinant strain in the gastrointestinal tract. Mice were immunised with an attenuated porcine Aujeszky's disease virus (ADV) vaccine. Serum antibody levels determined using a mouse Aujeszky's disease IgG ELISA showed that IgG levels were significantly higher in the pPG612.1-pLFN/L. pentosus group than in the PBS and Lactobacillus pentosus groups at days 7 and 21 (P < 0.01) and at day 14 (P < 0.05), indicating that this oral recombinant strain can improve the effectiveness of the vaccine and play a role in immune enhancement through humoral immunity. These results suggest that the recombinant Lactobacillus pentosus not only has the beneficial characteristics of lactic acid bacteria but also produces biologically functional lactoferrin.

Authors : Xu Yigang, Zong Xiaolin, Han Bing, Li Yijing, Tang Lijie,

(7) Infectious agents identified in aborted swine fetuses in a high-density breeding area: a three-year study.[TOP]

Pubmed ID :27400956
Publication Date : //
Reproductive failure in sows is one of the most important factors affecting pig breeding. Many reproductive disorders are linked to both environmental factors and infectious agents. The goal of our study was to determine the presence of pathogens that are known to cause abortion, considering a set of conditioning factors, such as seasonality and pregnancy period. A large number of aborted fetuses (1,625 fetuses from 140 farms) from a high-density breeding area in northern Italy was analyzed for a period of 3 years. The pigs were diagnosed based on direct (culture, PCR) or indirect (enzyme-linked immunosorbent assay) evidence. An infectious etiologic agent was found in 323 of 549 cases of abortion (58.8%). These included viral agents (Porcine circovirus-2, 138/323; Porcine reproductive and respiratory syndrome virus, 108/323; porcine parvovirus, 20/323; pseudorabies virus, 6/323; and Encephalomyocarditis virus, 3/323) and bacteria (Escherichia coli, 64/323; Streptococcus sp., 63/323; Staphylococcus sp., 5/323; Pasteurella sp., 3/323; Shigella sp., 1/323; and Yersinia sp., 1/323). This study describes the prevalence of infectious agents involved in reproductive failure in a high-density swine population. The data can be useful to swine breeders, practitioners, and medical specialists in monitoring animal health and in supervising the breeding process.

Authors : Salogni Cristian, Lazzaro Massimiliano, Giacomini Enrico, Giovannini Stefano, Zanoni Mariagrazia, Giuliani Matteo, Ruggeri Jessica, Pozzi Paolo, Pasquali Paolo, Boniotti Maria Beatrice, Alborali Giovanni Loris,

(8) [Immune Response of Recombinant Pseudorabies Virus rPRV-VP2 Expressing VP2 Gene of Porcine Parvovirus in Mice].[TOP]

Pubmed ID :27396164
Publication Date : //
In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.

Authors : Fu Pengfei, Pan Xinlong, Han Qiao, Yang Xingwu, Zhu Qianlei, Guo Xiaoqing, Zhang Yu, Chen Hongying,

(9) [Immunogenicity of a recombinant pseudorabies virus coexpressing ORF2 gene of PCV2 and porcine IL-18 gene in mice].[TOP]

Pubmed ID :27305786
Publication Date : //
To develop a bivalent vaccine against pseudorabies virus (PRV) and porcine circovirus (PCV2), IL-18 was used as immunologic adjuvant.

Authors : Guo Xiaoqing, Zhu Qianlei, Pan Xinlong, Yang Xingwu, Qiao Han, Wang Shujuan, Chen Hongying,

(10) A live, attenuated pseudorabies virus strain JS-2012 deleted for gE/gI protects against both classical and emerging strains.[TOP]

Pubmed ID :26946112
Publication Date : //
Emerging pseudorabies virus (PRV) variant have led to pseudorabies outbreaks in Chinese pig farms. The commercially available PRV vaccine provides poor protection against the PRV variant. In this study, a gE/gI deleted PRV strain JS-2012-△gE/gI was generated from a PRV variant strain using homologous DNA recombination. Compared to the parental strain JS-2012, JS-2012-△gE/gI grew slowly and showed small plaque morphology on Vero cells. The safety and immunological efficacy of JS-2012-△gE/gI was evaluated as a vaccine candidate. JS-2012-△gE/gI was avirulent to suckling piglets, but was able to provide full protection for young piglets against challenge with both the classical virulent PRV and the emerging PRV variant. After sows were vaccinated with the gE/gI-deleted strain, their suckling offspring were resistant to an otherwise lethal challenge with the classical and the variant PRVs. Piglets inoculated with JS-2012-△gE/gI did not develop PRV-specific gE-ELISA antibodies. Thus, JS-2012-△gE/gI appears to be a promising marker vaccine candidate to control PRV variant circulating in pig farms in China.

Authors : Tong Wu, Li Guoxin, Liang Chao, Liu Fei, Tian Qing, Cao Yanyun, Li Lin, Zheng Xuchen, Zheng Hao, Tong Guangzhi,