scholarly journals STANDARDIZATION OF MULTIPLEX REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION AND TYPING OF FOOT-AND-MOUTH DISEASE VIRUS PREVALENT IN BANGLADESH

2012 ◽  
Vol 8 (2) ◽  
pp. 149-155 ◽  
Author(s):  
MA Zinnah ◽  
MT Islam ◽  
MM Rahman ◽  
MT Hossain ◽  
MA Zinnah ◽  
...  

Foot-and-mouth disease (FMD) is a devastating viral disease of cattle that causes severe economic losses in terms of loss of production and calf mortality in Bangladesh. Despite of regular vaccination, outbreak of the disease has become a regular event throughout the country every year. Determination of prevailing serotypes of the causal agent foot-and-mouth disease virus (FMDV) is now crucial need for strategic vaccination programme. The present research work was aimed to standardize a multiplex RT-PCR assay typing of foot-and-mouth disease virus serotypes prevalent among cattle population of Bangladesh. Uniplex and multiplex RT-PCRs were successfully developed and standardized using the extracted RNA of reference FMDV (Type A, O and Asia 1) following adjustment of the concentration of the viral RNA of each serotype, volume of reaction mixture and thermal profile. The mPCR was evaluated on 82 field samples (vesicular fluid, tongue epithelium and tissue from inter-digital space) of the years 2007 and 2008. Of the 82 field samples, 56 (68.29%) were found positive for FMDV. The mPCR successfully differentiated single as well as dual serotypes infection. The serotypes A, O and Asia 1 were confirmed in the samples of the year 2007 and only serotype O in samples of the year 2008. Higher detection rate was found in vesicular fluid (100%) followed by tongue epithelium (79.66%). It may be concluded that the MRT-PCR standardized in this study could be used for detection and differentiation of FMDV serotypes using field samples.DOI = http://dx.doi.org/10.3329/bjvm.v8i2.11199 Bangl. J. Vet. Med. (2010). 8 (2) : 149-155 

2009 ◽  
Vol 16 (11) ◽  
pp. 1660-1664 ◽  
Author(s):  
Jae Ku Oem ◽  
Nigel P. Ferris ◽  
Kwang-Nyeong Lee ◽  
Yi-Seok Joo ◽  
Bang-Hun Hyun ◽  
...  

ABSTRACT A simple lateral-flow assay (LFA) based on a monoclonal antibody (MAb 70-17) was developed for the detection of foot-and-mouth disease virus (FMDV) under nonlaboratory conditions. The LFA was evaluated with epithelial suspensions (n = 704) prepared from current and historical field samples which had been submitted to the Pirbright Laboratory (United Kingdom) and from negative samples (n = 100) collected from naïve animals in Korea. Four FMDV serotypes (type O, A, Asia 1, and C) were detected in the LFA, but not the remaining three FMDV serotypes (SAT 1, SAT 2, and SAT 3). The diagnostic sensitivity of the LFA for FMDV types O, A, C, and Asia 1 was similar, at approximately 87.3%, to that of 87.7% obtained with antigen enzyme-linked immunosorbent assay (Ag-ELISA). The diagnostic specificity of the LFA was 98.8%, compared to 100% for the Ag-ELISA. These results demonstrate that the LFA using the FMDV MAb 70-17 to detect FMDV is a supportive method for taking rapid measurements at the site of a suspected foot-and-mouth disease outbreak in Asia before diagnosing the disease in the laboratory, thereby offering the possibility of implementing control procedures more rapidly.


2018 ◽  
Vol 6 (2) ◽  
pp. 23-26
Author(s):  
Mohammad Showkat Mahmud ◽  
Eusha Islam ◽  
Md. Giasuddin ◽  
Mohammed Abdus Samad ◽  
Md. Rezaul Karim ◽  
...  

2020 ◽  
Vol 2 (7A) ◽  
Author(s):  
Soumendu Chakravarti ◽  
Caroline Wright ◽  
Emma Howes ◽  
Richard Kock ◽  
Terry Jackson ◽  
...  

The picornavirus foot-and-mouth disease virus (FMDV) is responsible for one of the most significant diseases of livestock, leading to large economic losses due to reduced productivity and trade embargoes for areas not certified as disease-free. The picornavirus non-structural protein 3A is involved in replication of the viral RNA genome and is implicated in host tropism of several picornaviruses. Deletions in the C-terminus of 3A have been observed in FMDV outbreaks specific for swine and such viruses are non-pathogenic in cattle. The mechanism for species specific attenuation of FMDV is unknown. We have shown that FMDV containing a C-terminal deletion in 3A is attenuated in bovine cell culture and that the attenuated phenotype can be reversed by the JAK1/2 inhibitor Ruxolitinib (Rux), identifying a role for the induction of interferon stimulated genes (ISGs) in the restricted bovine tropism of the 3A-deleted virus.


2018 ◽  
Vol 30 (5) ◽  
pp. 699-707 ◽  
Author(s):  
Chungwon J. Chung ◽  
Alfonso Clavijo ◽  
Mangkey A. Bounpheng ◽  
Sabena Uddowla ◽  
Abu Sayed ◽  
...  

The highly contagious foot-and-mouth disease virus (FMDV) afflicts cloven-hoofed animals, resulting in significant costs because of loss of trade and recovery from disease. We developed a sensitive, specific, and rapid competitive ELISA (cELISA) to detect serum antibodies to FMDV. The cELISA utilized a monoclonal blocking antibody specific for a highly conserved FMDV nonstructural 3B epitope, a recombinant mutant FMDV 3ABC coating protein, and optimized format variables including serum incubation for 90 min at 20–25°C. Samples from 16 animals experimentally infected with one FMDV serotype (A, O, Asia, or SAT-1) demonstrated early detection capacity beginning 7 d post-inoculation. All samples from 55 vesicular stomatitis virus antibody-positive cattle and 44 samples from cloven-hoofed animals affected by non-FMD vesicular diseases were negative in the cELISA, demonstrating 100% analytical specificity. The diagnostic sensitivity was 100% against sera from 128 cattle infected with isolates of all FMDV serotypes, emphasizing serotype-agnostic results. Diagnostic specificities of U.S. cattle ( n = 1135) and swine ( n = 207) sera were 99.4% and 100%, respectively. High repeatability and reproducibility were demonstrated with 3.1% coefficient of variation in percent inhibition data and 100% agreement using 2 kit lots and 400 negative control serum samples, with no difference between bench and biosafety cabinet operation. Negative results from vaccinated, uninfected cattle, pig, and sheep sera confirmed the DIVA (differentiate infected from vaccinated animals) capability. This rapid (<3 h), select agent–free assay with high sensitivity and specificity, DIVA capability, and room temperature processing capability will serve as a useful tool in FMDV surveillance, emergency preparedness, response, and outbreak recovery programs.


Intervirology ◽  
2015 ◽  
Vol 58 (3) ◽  
pp. 190-196 ◽  
Author(s):  
Farahnaz Motamedi-Sedeh ◽  
Hoorieh Soleimanjahi ◽  
Amir Reza Jalilian ◽  
Homayoon Mahravani ◽  
Kamalodin Shafaee ◽  
...  

Objectives: Foot-and-mouth disease virus (FMDV) causes a highly contagious disease in cloven-hoofed animals and is the most damaging disease of livestock worldwide, leading to great economic losses. The aim of this research was the inactivation of FMDV type O/IRN/1/2007 to produce a gamma ray-irradiated (GRI) vaccine in order to immunize mice and guinea pigs. Methods: In this research, the Iranian isolated FMDV type O/IRN/1/2007 was irradiated by gamma ray to prepare an inactivated whole virus antigen and formulated as a GRI vaccine with unaltered antigenic characteristics. Immune responses against this vaccine were evaluated on mice and guinea pigs. Results: The comparison of the immune responses between the GRI vaccine and conventional vaccine did not show any significant difference in neutralizing antibody titer, memory spleen T lymphocytes or IFN-γ, IL-4, IL-2 and IL-10 concentrations (p > 0.05). In contrast, there were significant differences in all of the evaluated immune factors between the two vaccinated groups of mice and negative control mice (p < 0.05). The protective dose 50 for the conventional and GRI vaccines obtained were 6.28 and 7.07, respectively, which indicated the high potency of both vaccines. Conclusion: GRI vaccine is suitable for both routine vaccination and control of FMDV in emergency outbreaks.


2012 ◽  
Vol 93 (7) ◽  
pp. 1442-1448 ◽  
Author(s):  
Haiwei Wang ◽  
Mei Xue ◽  
Decheng Yang ◽  
Guohui Zhou ◽  
Donglai Wu ◽  
...  

Previously, we finely mapped the neutralizing epitopes recognized by foot-and-mouth disease virus (FMDV) type Asia1-specific mAb 3E11 and FMDV type O-specific mAb 8E8. In this study, we engineered recombinant FMDVs of the serotype Asia1 (rFMDVs) displaying the type O-neutralizing epitope recognized by the mAb 8E8. These epitope-inserted viruses were genetically stable and exhibited growth properties that were similar to those of their parental virus. Importantly, the recombinant virus rFMDV-C showed neutralization sensitivity to both FMDV type Asia1 and type O mAbs, as well as to polyclonal antibodies. These results indicated that this epitope-inserted virus has the potential to induce neutralizing antibodies against both FMDV type Asia1 and type O. Our results demonstrated that the G-H loop of FMDV type Asia1 effectively displays the protective neutralizing epitopes of other FMDV serotypes, making this an attractive approach for the design of novel FMDV vaccines.


2008 ◽  
Vol 82 (18) ◽  
pp. 9075-9085 ◽  
Author(s):  
Vivian O'Donnell ◽  
Michael LaRocco ◽  
Barry Baxt

ABSTRACT Foot-and-mouth disease virus (FMDV) utilizes different cell surface macromolecules to facilitate infection of cultured cells. Virus, which is virulent for susceptible animals, infects cells via four members of the αV subclass of cellular integrins. In contrast, tissue culture adaptation of some FMDV serotypes results in the loss of viral virulence in the animal, accompanied by the loss of virus' ability to use integrins as receptors. These avirulent viral variants acquire positively charged amino acids on surface-exposed structural proteins, resulting in the utilization of cell surface heparan sulfate (HS) molecules as receptors. We have recently shown that FMDV serotypes utilizing integrin receptors enter cells via a clathrin-mediated mechanism into early endosomes. Acidification within the endosome results in a breakdown of the viral capsid, releasing the RNA, which enters the cytoplasm by a still undefined mechanism. Since there is evidence that HS internalizes bound ligands via a caveola-mediated mechanism, it was of interest to analyze the entry of FMDV by cell-surface HS. Using a genetically engineered variant of type O1Campos (O1C3056R) which can utilize both integrins and HS as receptors and a second variant (O1C3056R-KGE) which can utilize only HS as a receptor, we followed viral entry using confocal microscopy. After virus bound to cells at 4°C, followed by a temperature shift to 37°C, type O1C3056R-KGE colocalized with caveolin-1, while O1C3056R colocalized with both clathrin and caveolin-1. Compounds which either disrupt or inhibit the formation of lipid rafts inhibited the replication of O1C3056R-KGE. Furthermore, a caveolin-1 knockdown by RNA interference also considerably reduced the efficiency of O1C3056R-KGE infection. These results indicate that HS-binding FMDV enters the cells via the caveola-mediated endocytosis pathway and that caveolae can associate and traffic with endosomes. In addition, these results further suggest that the route of FMDV entry into cells is a function solely of the viral receptor.


PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4823 ◽  
Author(s):  
Guoqiang Wang ◽  
Yunchao Liu ◽  
Hua Feng ◽  
Yumei Chen ◽  
Suzhen Yang ◽  
...  

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals that has caused tremendous economic losses worldwide. In this study, we designed a chimeric nanoparticles (CNPs) vaccine that displays the predominant epitope of the serotype O foot-and-mouth disease virus (FMDV) VP1 131-160 on the surface of MS2 phage. The recombinant protein was expressed inEscherichia Coliand can self-assemble into CNPs with diameter at 25–30 nmin vitro. A tandem repeat peptide epitopes (TRE) was prepared as control. Mice were immunized with CNPs, TRE and commercialized synthetic peptide vaccines (PepVac), respectively. The ELISA results showed that CNPs stimulated a little higher specific antibody levels to PepVac, but was significantly higher than the TRE groups. Moreover, the results from specific IFN-γ responses and lymphocyte proliferation test indicated that CNP immunized mice exhibited significantly enhanced cellular immune response compared to TRE. These results suggested that the CNPs constructed in current study could be a potential alternative vaccine in future FMDV control.


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