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1.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(1):10-19, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-2056573

ABSTRACT

The aim of this study is to establish an indirect ELISA technique for detecting the SIgA antibody against porcine epidemic diarrhea virus (PEDV) to evaluate its mucosal immunity. Firstly, the S1D gene (534-789 aa) of PEDV was cloned into the pET-28a(+) vector, and induced in Escherichia coli BL21 (DE3) by IPTG, the product of which was in the form of inclusion bodies. According to Western-blot, the target protein S1D with antigenic activity was 32 ku in molecular weight and could be well detected. Then, the S1D protein was denatured by 8 mol/L urea, purified and gradient as the coating antigen to establish an indirect ELISA for detecting the PEDV specific SIgA antibody in nasal or oral mucus by optimizing conditions. And the optimal antigen coating concentration of ELISA was 2 micro g mL, the working concentrations of nasal mucus was 1:1 and the optimal blocking solution was 50 g/L skimmed milk, while the working concentrations and optimal blocking solution were 1:2 and 30 g/L BSA in oral mucus, the working concentrations of the enzyme-labeled antibody was 1:2 000 in nasal and oral mucus. Finally, 84 samples of oral and nasal mucus from immunized pigs were detected by S1D of ELISA, and the coincidence rate could reach 95.2% compared with purified PEDV of ELISA. In conclusion, the indirect ELISA established in this study provided a quick, simple, sensitive, and specific method to detect PEDV specific SigA for evaluating the level of PEDV mucosal immunity.

2.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(12):1500-1508, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-2040500

ABSTRACT

Based on the M gene sequence of TGEV and PEDV and VP2 gene sequence of PoRV, the optimal reaction system and amplification procedure were established by optimizing primer, probe concentration and annealing temperature, and the Quantitative PCR method of TaqMan probes for three viruses is successfully established. On this basis, after further optimization of conditions, a triple real-time fluorescent quantitative PCR method for detecting TGEV, PEDV, and PoRV was established. The detection sensitivity of this method for TGEV, PEDV, and PoRV were 2.49 copies/ L, 4.36 copies/ L, and 4.96 copies/ L respectively. The maximum value of CV in repeated trials detected by TGEV, PEDV and PoRV were 2.5%, 3.8%, 4.3%, and the maximum value of CV in repeated trials between groups were 3.7%, 3.4%, 3.2%, which are no more than 5%.indicating that the established method has good reproducibility. Using this method to detect PRV, PCV1, and PRRSV virus samples, there is no cross-reaction, indicating that the method is specific. Using the established method to detect 40 clinical diseases, the samples were tested, and the positive rates of TGEV, PEDV, and PoRV were 5%, 30%, and 12.5%respectively. The mixed infection rate of TGEV and PEDV was 2.5%, the mixed infection rate of PEDV and PoRV was 5%. The results of the multiple fluorescence quantitative PCR method are consistent with those of the detection of a single fluorescent RT-PCR method, indicating that the established method has good clinical application value.

3.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(11):1373-1378, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-2040499

ABSTRACT

In order to build a specific, sensitive and rapid detection method for PAstV3 detection, the PAstVB gene sequences in Genbank were used and the conserved region in ORFlb was selected to design specific primers and TaqMan probe. Clinical stool samples were collected and preliminary detected by this newly established real-time RT-PCR method after reaction systems and conditions optimization. This detection method established in this study has a good linear relationship with the standard curve, with R2 value up to 0.9971. The sensitivity is 100 times higher than conventional PCR method, The variation co-efficient of in-batch and inter-batch repeatability test is less than 2.0%, indicating good repeatability. The detection results of Clinical samples showed that the positive rate of this method is higher than conventional PCR method. The establishment of this method provides a rapid detection means for PAstV3 laboratory diagnosis and epidemiological investigation.

4.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(11):1421-1427, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-2040498

ABSTRACT

Recently, the variation and isolation of porcine epidemic diarrhea Virus (PEDV) has been a focus of industry research. Whether porcine aminopeptidase (pAPN) is a functional receptor of PEDV infection is still controversial. Therefore, this article aims to review the latest progress on pAPN as a receptor of PEDV and its role during infection, to clarify whether pAPN is a functional receptor and to provide a reference for isolation and subsequent study of PEDV.

5.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(11):1341-1347, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-2040497

ABSTRACT

The recombinant expression plasmid pIRES-S1 was constructed according to the gene sequence of PEDV S1 in NCBI (GenBank:JQ517274). The plasmid pIRES-S1 was transfected into ST cells by electrotransfer. After G418 pressurization screening, western-blot detection and suspension domestication, a stable transduction cell pool expressing S1 protein was obtained. The results of Western-blot showed that S1 protein have good reactivity. An indirect ELISA was established by using S1 protein as coating antigen, and the ELISA was used to detect PEDV clinical serum and PEDV negative serum of imported breeder pigs. Take the serum neutralization test as the standard, the results showed that the sensitivity of the ELISA was 96.3% and the specificity was 97.7%.It was significantly consistent with the serum neutralization test (kappa value=0.882, P < 0.05). The ELISA was used to detect the tracking serum of PEDV back-feeding pigs. The results showed that it could accurately evaluate the growth and decline of PEDV Ig G antibody level in infected pigs. Our results suggested that the ELISA based on S1 protein established in this study has high sensitivity and specificity. It could be used to detect PEDV antibody in clinical serum samples and provide an effective basis for immune evaluation of PEDV in pigs.

6.
Acta Microbiologica Sinica ; 8:3152-3165, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-2040441

ABSTRACT

Objective: To identify the key host protein that can regulate the replication of porcine epidemic diarrhea virus (PEDV).

7.
Journal of South China Agricultural University ; 41(5):27-35, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-2040361

ABSTRACT

Objective: To prepare monoclonal antibodies against porcine epidemic diarrhea virus (PEDV) N protein, and develop an indirect immuno-fluorescence assay method used for detecting PEDV. Method: The expressed recombinantly PEDV N protein was used as an immunogen and 8-week-old female BALB/c mice were immunized. Then their spleen cells with high antibody titer were isolated and fused with SP2/0 cells. The hybridoma cell lines secreting monoclonal antibodies against PEDV N protein were screened. In Vero cells infected with PEDV, monoclonal antibody of anti-PEDV N protein was used as the primary antibody and FITC-goat-anti-mouse IgG was used as the secondary antibody to develop indirect immuno-fluorescence assay method used for detecting PEDV. Result: The prepared hybridoma cell lines could stably secrete anti-PEDV N protein antibodies, ELISA antibody titer in cell supernatant was above 1:3 200, and in mouse ascites above 1:1 000 000. While monoclonal antibodies were applied in established indirect immuno-fluorescence assay, the optimal conditions were that cells were fixed with 80% () acetone at -20 degrees C for 30 min;The primary antibody was diluted 1 000 times by PBS buffer solution and incubated at 4 degrees C overnight;The secondary antibody was diluted 100 times by PBS buffer solution and incubated at 37 degrees C for 1 h. Transmissible gastroenteritis virus (TGEV), classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine reproductive virus (PRV), porcine enteric a corone virus (PEAV), porcine rotavirus (PoRV) and PEDV were detected by established indirect immuno-fluorescence assay method, only PEDV showed positive, all the else viruses showed negative.

8.
Acta Veterinaria et Zootechnica Sinica ; 53(7):2260-2267, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-2025546

ABSTRACT

The C-terminal domain (CTD) of porcine deltacoronavirus S1 subunit is the main region which induces the neutralizing antibody. S1-CTD was expressed by HEK-293T eukaryotic expression system and purified, and porcine ileal epithelium cells membrane proteins were extracted to investigate porcine host proteins that interact with it. Thirty-two suspected interacting host proteins were obtained by co-inmunprecipitation (Co-IP) and mass spectrometry. Eukaryotic expression plasmid of KIF1 binding protein (KIFBP) was constructed, and the interaction between KIFBP and S1-CTD was identified by Co-IP and laser confocal microscopy. All results proved that KIFBP interacted with S1-CTD and co-located in cytoplasm. Further research indicated that overexpression of KIFBP could effectively reduce the viral mRNA level and the viral titer in which the mRNA level decreased by about 70%, and the viral titer decreased by 101.6TCID50. In conclusion, a host protein KIFBP interacting with PDCoV S1-CTD was screened and identified in this study which provides a theoretical basis for understanding the pathogenesis of PDCoV.

9.
Acta Veterinaria et Zootechnica Sinica ; 53(6):2024-2028, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-2025545

ABSTRACT

This study aimed to analyze the proliferation characteristics of porcine deltacoronavirus (PDCoV) in suspension cultured porcine kidney cells LLC-PK1, so as to provide Candidate cell for large-scale production of PDCoV inactivated vaccine. LLC-PK1 cells were suspended by gradually decreasing serum method. PDCoV adaptive monoclonal cell lines were screened by limited dilution method. Indirect immunofluorescence method was used to identify the infectivity of PDCoV. The initial cell density, MOI, time of receiving virus collection and TPCK pancreatin concentration were screened to determine the best suspension culture conditions. The suspension cell strain LLC-PK1Sa which can proliferate PDCoV efficiently was screened out;PDCoV can specifically infect LLC-PK1 cells;PDCoV inoculated LLC-PK1Sa cells with a density of 2 x 106 cells.mL-1 according to the MOI of 10-3, When the final concentration of TPCK pancreatin reached 7.5 g.mL-1, the titer of virus solution harvested 48 h after inoculation was the highest. In this study, the efficient proliferation of PDCoV in LLC-PK1Sa suspension cells was realized for the first time, and the suspension culture conditions were preliminarily optimized, which could provide theoretical reference for large-scale production of PDCoV inactivated vaccine.

10.
Zycie Weterynaryjne ; 95(9):554-559, 2020.
Article in Polish | CAB Abstracts | ID: covidwho-2011448

ABSTRACT

Throughout the course of civilization, epidemics and pandemics have ravaged humanity, destroyed animal breeding and horticulture, and has also changed the course of history. It has been estimated that Justinian plague has affected half of the population of Europe and killed in three pandemics 50 million people, the avian-borne flu (Spanish flu), resulted in 50 million deaths worldwide in the years 1918-1919, and recently the COVID-19 is officially a pandemic, after barreling through 114 countries in just three months. In the past, rinderpest has hit Europe with three long panzootics, African swine fever (ASF), is still a threat to both the swine production industry and the health of wild boar populations. Several molecular changes occur in the pathogen that may trigger an epidemic or even pandemic. These include increase of virulence, introduction into a novel host, and changes in host susceptibility to the pathogen. Once the infectious disease threat reaches an epidemic or pandemic level, the goal of the response is to mitigate its impact and reduce its incidence, morbidity and mortality as well as disruptions to economic, political, and social systems. An epidemic curve shows progression of illnesses in an outbreak over time and the SIR, SI, SIRD and SEIR represent the simplest compartmental models that enable simplify the mathematical modelling of epidemics. This article throws a light on changing ideas in epidemiology of infectious diseases.

11.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(5):603-609, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-1994656

ABSTRACT

To establish a J2-KD (knockdown) cell line stably expressing interfered IFITM1 and study the effect of interference with IFITMI gene on the infection of PCV2, PRV and TGEV. Gene cloning tech- niques were used to constructed pLKO. l-EGFP-Puro-IFITMI recombinant vector, which was co-transfected into 293 FT cells with lentiviral packaging plasmids psPAXZ and pMDZ. G to produce green fluorescent protein labeled lentiviruses expression IFITMlshRNA, the viral supernatant was collected at 48 hours after post transfection. J2 cells were infected with the harvested lentiviruses, screened by puromycin and cloned via cell limited dilution. Real-time PCR identify that the cell lines with stable interference with IFITMl gene were obtained, and via MTT method verify that interference with IFITMI expression had no effect on the growth of J2 cells, the successfully constructed J2 stable cell line interfere with IFITMl expression was named as JZ-KD. PRV, PCV2 and TGEV infected J2-KD cells, respectively. Using real-time fluorescence quantitative PCR detect virus replication. The results showed that J2-KD cell line was successfully generated with interfered IFITMl expression;the copy number of PCV2 and TGEV were in- creased, while PRV was decreased in J'Z-KD cell. Indicating that the interference of IFITMI gene expression markedly inhibited the replication of PRV while promoted that. of TGEV and PCV2, providing a basis for further study on the function of porcine IFITMI protein and elucidates its antiviral mechanism.

12.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(9):1147-1158, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-1994654

ABSTRACT

To understand the genetic diversity of porcine deltacoronavirus(PDCo V) in Guangxi Province, clinical diarrhea samples were collected from suspected piglets in Guangxi Province from2017 to 2019, detected by RT-PCR for PDCoV, and the positive samples were used for amplification and sequence of S, M, N genes. Finally, 16 S, M and N gene sequences of PDCoV were obtained. Homology analysis showed that the S, M, N gene nucleotide identity among Guangxi strains were 95.8% -99.9%, 95.9%-100% and 97.9%-99.9%, respectively. The nucleotide identity of S, M and N genes among Guangxi strains and other reference strains were 95.1%-100%, 95.0%-100%and 96.3%-99.9%, respectively. Sequence alignment showed that S1 protein existed amino acid mutations and insertions, and there were some variations among different epidemic strains. Phylogenetic trees based on S, M and N genes obtained similar topological diagram and all strains could be divided into Group I, Group II and GroupIII, of which Group I came from USA, Japan and Korea, Group II came from China, and Group III came from China, Vietnam, Laos and Thailand. Most strains from Guangxi Province distributed in Group II, individual strain distributed in Group III and some strains formed a single small branch. The evolutionary rates of S, M and N genes of Guangxi strains and other reference strains were 2.57 x 10-4, 2.07 x 10-4, 1.70 x 10-4 substitutions/site/year, respectively, showing that the evolutionary rate of S gene was the fastest. The results indicated that the S, M, N genes of PDCo V strains from Guangxi Province had some variations and existed genetic diversity.

13.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(5):537-544, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-1994651

ABSTRACT

Long noncoding RNA (lncRNA) is a type of non-coding RNA molecule longer than 200 nt, which plays vital roles in biological events. Our previous results demonstrated that the host's lncRNA expression profile was significantly changed after porcine epidemic diarrhea virus (PEDV) infection. In this study, one of the lncRNAs, lncRNA9606, was selected to investigate its impact on PEDV replication. First, the kinetics of lncRNA9606 expression in IPEC-J2 cells were examined at different time points after PEDV infection. The results confirmed that PEDV infection significantly upregulated the expression of lncRNA9606. The lncRNA9606 expression levels in different cells or tissues were evaluated and the results showed that the amount of lncRNA9606 in Peyer's patches and peripheral blood mononuclear cells were significantly higher than that in small intestinal epithelial cell lines. It was mainly localized in the nucleus. Further investigations indicated that over expression of lncRNA in LLC-PK1 cells significantly inhibited PEDV replication. In conclusion, lncRNA9606 can suppress the PEDV replication in LLC-PK1 cells.

14.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(5):556-562, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-1994650

ABSTRACT

In order to establish an efficient, sensitive and specific semi-nest RT-PCR method for the detection of Transmissible gastroenteritis virus (TGEV), three specific primers were designed according to the N gene published by GenBank, the reaction system was established and optimized, and specificity and sensitivity were detected. The results showed that the method could successfully amplify the bands of 483 bp and 338 bp, and had good specificity to TGEV, there is no cross reaction with PEDV, PRov, PBov and PDCov, and the lowest sensitivity was 1.86 x 10-1 pg/L. The semi-nest RT-PCR shown the positive rate was 36% in 50 samples of pig diarrhea, which was higher than that of common RT-PCR, and then the positive samples coincidence rate was 100%. This semi-nest RT-PCR method has high sensitivity and specificity, and can accurately diagnose TGEV infection, which provides an effective method for clinical detection and epidemiological investigation of TGEV.

15.
Acta Veterinaria et Zootechnica Sinica ; 53(5):1587-1597, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-1994513

ABSTRACT

HEK293 cells were used as the cell model to investigate the role of human aminopeptidase N (hAPN) in the invasion of porcine deltacoronavirus (PDCoV) into human cells. The proliferation of PDCoV on HEK293 cells was firstly identified by RT-qPCR/RT-PCR. And then, hAPN knockout cell line was constructed by CRISPR/Cas9 technology and cell viability of HEK293 hAPN knockout and wild-type cells was verified by CCK-8 assay. Effect of hAPN knockout and overexpression on PDCoV replication was detected by RT-qPCR and Western blot. Meanwhile, interaction of PDCoV S protein and hAPN protein was analyzed by homology modeling and molecular docking. Results showed that PDCoV virus copies rapidly increased at 12-36 h and reached peak level at 36 h, it could propagate at least for passage 2 on HEK293 cells. There was no significant difference in cell viability between hAPN knockout cells and wild-type cells. Knockout of hAPN inhibit PDCoV replication and overexpression of hAPN enhance PDCoV replication. Homology modeling and molecular docking analysis showed S1 protein could bind hAPN domain II. Residues TYR92, THR51, THR48, PHE16 and MET14of S1 protein receptor binding motif 1 (RBM1) can form hydrogen bonds with residues PHE490, GLN531, ARG528 and SER529 of hAPN. This study indicates that hAPN plays a critical role in HEK293 cells during PDCoV infection, which provides new theoretical evidence for further studies on the mechanism of PDCoV entry into host cells and cross-species transmission.

16.
Acta Veterinaria et Zootechnica Sinica ; 53(5):1536-1543, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-1994512

ABSTRACT

This study aims to investigate the protective effect of infected piglets which were immunized with different dose of inactivated porcine epidemic diarrhea virus (PEDV) vaccines. The number of infective virus particles and total virus particles of PEDV with different concentrations were determined, and the mice were immunized with different concentration vaccine prepared as antigen, respectively. The humoral and cellular immune production were determined by ELISA antibody detection method, neutralization test and ELISPOT method. Vaccine with appropriate antigen content was selected to immunize piglets, then the antibody was determined. The relationship between concentrated vaccine and protective effect was studied by challenge experiment. The results showed that, when the antigen dose was equal or greater than 8x106 pfu.mL-1, the inactive vaccine could effectively stimulate mice to produce humoral and cellular immunity. The piglets immunized with 2 mL inactivated PEDV vaccine containing 8x106 pfu.mL-1 antigen could resist diarrhea and continuous viral shedding caused by PEDV challenge. Compared with the total number of virus particles, the number of infectious virus particles was significantly correlated with antibody production (r=0.998 1), and neutralization titer was significantly correlated with piglet protection (r=0.974 7). PEDV inactivated vaccine can provide good immune protection, in which the number of infectious virus particles is the key factor to improve the antibody level. Antibody titer, as an index of humoral immunity, is an important reference for judging immune protection.

17.
Chinese Veterinary Science / Zhongguo Shouyi Kexue ; 50(6):671-678, 2020.
Article in Chinese | CAB Abstracts | ID: covidwho-1975502

ABSTRACT

In present study, in order to develop a new and effective porcine epidemic diarrhea virus (PEDV)vaccine, three B cell epitopes and the truncated S1 gene of PEDV spike protein were combined and inserted into the immunodominant region of the HBcAg. Then the constructed recombinant plasmid HBcAg-PE was transformed to E. coli BL21 (DE3) for expression. After purification and identification by Western-blot, the expressed recombinant proteins HBPE were injected into BALB/c mice as vaccine antigen with different doses through intramuscular injection and its immune effect were preliminary evaluated. The results showed that the recombinant proteins HBPE was expressed as precipitation form and it could reacted specifically with PEDV-positive serum after purification and renaturation. Besides, the RH could induce anti-PEDV specific antibodies and the related Thl and Th2 cytokines in mice. The above results indicate that the recombinant compound epitope antigen of PEDV was successfully constructed. and its immunogenicity as a new vaccine candidate was evaluated in the mice in this study. The results of this study provided a new idea for the development of PEDV genetic engineering vaccine in the future.

18.
Acta Veterinaria et Zootechnica Sinica ; 53(4):1173-1181, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-1975364

ABSTRACT

The purpose of this study is to establish a blocking ELISA antibodies detection method for porcine epidemic diarrhea virus (PEDV). The purified N protein was used as the coating antigen, and the ELISA reaction conditions were optimized by the chess rboard titration. A blocking ELISA method for detecting PEDV antibodies was established, and its specificity, sensitivity and repeatability tests were carried out. One hundred and forty clinical serum samples were tested, and the results were compared with commercially IDvet PEDV indirect ELISA antibodies detection kit. The results showed that the best antigen coating concentration was 625 ng.mL-1, and the best dilution ratio of serum was 1:1;The best dilution of the HRP-conjugated antibody working solution was 1:5 000;There was no cross-reaction with healthy pig serum and the positive sera of common pig disease pathogens, such as classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), and transmissible gastroenteritis virus (TGEV). The sensitivity of PEDV positive serum was 1:16, which was equivalent to that of IDvet ELISA kit (titer 1:32). The coefficient of variation of within-run and between-run repeatability test is less than 10%, so it showed that the blocking ELISA established in this study had good repeatability and stability;the kappa value of detected 140 clinical porcine serum using this method was 0.87 when compared with IDvet ELISA. The above results indicated that the established blocking ELISA method for detecting PEDV antibodies in this study could be applied to the prevention and control of PEDV, epidemiological investigation and the monitoring of antibody levels after vaccine immunization.

19.
Acta Agriculturae Jiangxi ; 34(2):160-165, 2022.
Article in Chinese | CAB Abstracts | ID: covidwho-1964892

ABSTRACT

In this study, 650 tissue samples which were collected from 16 pig farms in Hubei Province, were used to detect porcine circovirus (PCV) and Porcine epidemic diarrhea virus (PEDV). The results showed that the positive rates of PCV1, PCV2, PCV3 and PEDV single infection were 1.08%, 4.15%, 2.46% and 6.46%, respectively. In the double infections, PEDV+PCV2 had the highest positive rate of 3.54%, followed by PCV2+PCV3, with a positive rate of 1.54%. In multiple infections, PEDV+PCV2+PCV3 had the highest positive rate of 2.00%. The results indicated that the positive rates of PEDV and PCV were decreased compared with the previous studies, but the prevalence of PEDV and PCV was still wide in Hubei Province, and most of which were co-infection.

20.
Journal of Yangzhou University, Agricultural and Life Sciences Edition ; 42(6):48-53, 2021.
Article in English, Chinese | CAB Abstracts | ID: covidwho-1964809

ABSTRACT

As a member of the family Picornaviridae, porcine sapelovirus (PSV) is often infected with porcine epidemic diarrhea virus, teschovirus and so on. In recent years, PSV has been isolated from porcine in many provinces of China. It suggests that it is necessary to strengthen the research on PSV. In this study, according to the sequence of PSV HuN2 strain, VP1 gene was inserted into the pGEX-6 P-1 vector, and expressed the recombinant protein. BALB/c mice aged 6-8 weeks were immunized according to the standard procedure. After the third immunization, the mouse orbital blood was collected to identify the antibody level. The highly positive mouse spleen cells were selected for cell fusion. The positive hybridoma cells and two subclones were screened by IFA method, and then a PSV VP1 monoclonal antibody was obtained, named as 33-2 A. The results of IFA showed that PSV could be recognized by 33-2 A MAb, and specific green fluorescence appeared in the cytoplasm;The results of WB and IP showed that PSV infected porcine cell could specifically bind to 33-2 A, and there was a specific band at 32 ku. We also identified the B-cell antigen epitope of 33-2 A, it was at amino acids 40-46 of PSV VP1 protein, and the polypeptide sequence was 40PALTAAE46. The results showed that the monoclonal antibody can react with PSV VP1 protein. The epitope was analyzed with the PSV sequences uploaded in NCBI, 33-2 A antibody can react with most PSV strains and has a certain universal to PSV. This study laid a foundation for the study of the etiology and pathogenesis of PSV.

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