scholarly journals Isolation, Serotyping and Molecular Detection of Bovine FMD Virus from Outbreak Cases in Aba'ala District of Afar Region

2020 ◽  
Author(s):  
Teshager Dubie Tegegne ◽  
Tsedale Amare Mengiste
Keyword(s):  
Molecular Detection ◽  
Vaccine Development ◽  
Universal Primers ◽  
Clinical Samples ◽  
Effective Vaccine ◽  
Rt Pcr ◽  
Documentation System ◽  
Fmd Virus ◽  
Afar Region ◽  
Fmdv Serotypes

Abstract Background: Among the top listed economically important transboundary livestock diseases of cattle, foot and mouth disease (FMD) is the leading bottleneck in livestock production and productivity in Ethiopia. On the basis of FMDV active outbreak cases, a cross sectional study was undertaken to collect samples from January, 2019 to March, 2020 intended for isolation, serotyping and molecular detection of FMDV in the study district. Purposive sampling method was applied to select the study area for the reason that the presence of current active FMD outbreak case report during the study period. Totally, 27 FMD suspected clinical samples were collected from clinically affected study population during field outbreak. Out of 27 samples, 18 of them were inoculated on cultured Baby hamster kidney (BHK-21) monolayer cells and all 27 samples were tested using conventional RT-PCR and sets of specific universal primers. Finally, the PCR products were visualized with UV illumination and imaged with gel documentation system. Results: The current study result revealed that out of 18 clinical samples subjected to virus isolation, 72.2%(n=13) of these cultures exhibited FMDV induced cytopathic effect (CPE) and the identified serotype was SAT-2 FMD virus. Out of 27 clinical samples tested by conventional RT-PCR, only 12 FMDV samples were found to be FMDV positive by universal primers. Out of 27 clinical samples detected by conventional reverse transcription polymerase chain reaction (RT-PCR), only 12 FMDV samples were found to be FMDV positive by universal primers.Conclusions: Our study finding indicated FMDV is prevalent in the study area and FMDV serotype SAT-2 was the causality for the outbreaks of the disease in the study area. Hence, region wise regular FMD outbreaks investigation, further phylogenetic analysis and vaccine matching field isolates should be carried out to know in depth data about FMDV serotypes and topotypes involving in afar region of Ethiopia for effective vaccine development and control of the disease.

scholarly journals Isolation, Serotyping and Molecular Detection of Bovine FMD Virus from Outbreak Cases in Aba'ala District of Afar Region

2020 ◽  
Author(s):  
Teshager Dubie Tegegne ◽  
Tsedale Amare Mengiste
Keyword(s):  
Molecular Detection ◽  
Vaccine Development ◽  
Universal Primers ◽  
Clinical Samples ◽  
Effective Vaccine ◽  
Rt Pcr ◽  
Documentation System ◽  
Fmd Virus ◽  
Afar Region ◽  
Fmdv Serotypes

Abstract Background: Among the top listed economically important transboundary livestock diseases of cattle, foot and mouth disease (FMD) is the leading bottleneck in livestock production and productivity in Ethiopia. On the basis of FMDV active outbreak cases, a cross sectional study was undertaken to collect samples from January, 2019 to March, 2020 intended for isolation, serotyping and molecular detection of FMDV in the study district. Purposive sampling method was applied to select the study area for the reason that the presence of current active FMD outbreak case report during the study period. Totally, 27 FMD suspected clinical samples were collected from clinically affected study population during field outbreak. Out of 27 samples, 18 of them were inoculated on cultured Baby hamster kidney (BHK-21) monolayer cells and all 27 samples were tested using conventional RT-PCR and sets of specific universal primers. Finally, the PCR products were visualized with UV illumination and imaged with gel documentation system. Results: The current study result revealed that out of 18 clinical samples subjected to virus isolation, 72.2%(n=13) of these cultures exhibited FMDV induced cytopathic effect (CPE) and the identified serotype was SAT-2 FMD virus. Out of 27 clinical samples tested by conventional RT-PCR, only 12 FMDV samples were found to be FMDV positive by universal primers. Out of 27 clinical samples detected by conventional reverse transcription polymerase chain reaction (RT-PCR), only 12 FMDV samples were found to be FMDV positive by universal primers.Conclusions: Our study finding indicated FMDV is prevalent in the study area and FMDV serotype SAT-2 was the causality for the outbreaks of the disease in the study area. Hence, region wise regular FMD outbreaks investigation, further phylogenetic analysis and vaccine matching field isolates should be carried out to know in depth data about FMDV serotypes and topotypes involving in afar region of Ethiopia for effective vaccine development and control of the disease.

scholarly journals Isolation, Serotyping, and Molecular Detection of Bovine FMD Virus from Outbreak Cases in Abaʼala District of Afar Region, Ethiopia

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Teshager Dubie ◽  
Tsedale Amare
Keyword(s):  
Molecular Detection ◽  
Vaccine Development ◽  
Universal Primers ◽  
Clinical Samples ◽  
Effective Vaccine ◽  
Rt Pcr ◽  
Documentation System ◽  
Cross Sectional ◽  
Fmd Virus ◽  
Study Results

Background. On the basis of FMDV outbreak cases, a cross-sectional study was undertaken to collect samples from January 2019 to March 2020 intended for isolation, serotyping, and molecular detection of FMDV in the study district. The purposive sampling method was applied to select the study area for the reason of the presence of FMD outbreak case report during the study period. Totally, 27 FMD clinical samples were collected from affected study population during field outbreak. Out of 27 samples, 18 of them were inoculated on cultured Baby hamster kidney (BHK-21) monolayer cells, and all 27 samples were tested using conventional RT-PCR and sets of specific universal primers. Finally, the PCR products were visualized with UV illumination and imaged with gel documentation system. Results. The current study results revealed that out of 18 clinical samples subjected to virus isolation, 72.2% (n = 13) of these cultures exhibited FMDV-induced cytopathic effect (CPE) and the identified serotype was SAT-2 FMD virus. Out of 27 clinical samples tested by conventional RT-PCR, only 12 FMDV samples were found to be FMDV positive by universal primers. Out of 27 samples detected by conventional RT-PCR, only 12 FMDV samples were found to be FMDV positive by universal primers. Conclusions. Our study finding indicated that FMDV is prevalent in the study area and FMDV serotype SAT-2 was the causality for the outbreaks of the disease in the study area. Hence, region-wise FMD outbreak investigation, further phylogenetic analysis, and vaccine matching field isolates should be carried out for effective vaccine development to control the disease.

scholarly journals High speed large scale automated isolation of SARS-CoV-2 from clinical samples using miniaturized co-culture coupled with high content screening

2020 ◽  
Author(s):  
Rania Francis ◽  
Marion Le Bideau ◽  
Priscilla Jardot ◽  
Clio Grimaldier ◽  
Didier Raoult ◽  
...  
Keyword(s):  
High Speed ◽  
Large Scale ◽  
Vaccine Development ◽  
Work Load ◽  
Drug Susceptibility Testing ◽  
High Content Screening ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Positive Elements

AbstractSARS-CoV-2, a novel coronavirus infecting humans, is responsible for the current COVID-19 global pandemic. If several strains could be isolated worldwide, especially for in-vitro drug susceptibility testing and vaccine development, few laboratories routinely isolate SARS-CoV-2. This is due to the fact that the current co-culture strategy is highly time consuming and requires working in a biosafety level 3 laboratory. In this work, we present a new strategy based on high content screening automated microscopy (HCS) allowing large scale isolation of SARS-CoV-2 from clinical samples in 1 week. A randomized panel of 104 samples, including 72 tested positive by RT-PCR and 32 tested negative, were processed with our HCS procedure and were compared to the classical isolation procedure. Isolation rate was 43 % with both strategies on RT-PCR positive samples, and was correlated with the initial RNA viral load in the samples, where we obtained a positivity threshold of 27 Ct. Co-culture delays were shorter with HCS strategy, where 80 % of the positive samples were recovered by the third day of co-culture, as compared to only 25 % with the classic strategy. Moreover, only the HCS strategy allowed us to recover all the positive elements after 1 week of co-culture. This system allows rapid and automated screening of clinical samples with minimal operator work load, thus reducing the risks of contamination.

scholarly journals Molecular characterization of foot-and-mouth disease viruses collected from Northern and Central Ethiopia during the 2018 outbreak

2020 ◽  
Vol 13 (3) ◽  
pp. 542-548
Author(s):  
Yeneneh Tesfaye ◽  
Fazlurrahman Khan ◽  
Esayas Gelaye
Keyword(s):  
Gene Sequencing ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Serotype A ◽  
Serotype O ◽  
Fmdv Serotypes ◽  
Virus Serotype ◽  
Foot And Mouth

Background and Aim: Foot-and-mouth disease (FMD) is endemic in several developing countries and affects poor farmers through loss of production, death of diseased animals, and loss of animal byproducts. Forty-three samples were collected from 12 sites of five geographical located areas from suspected FMD virus (FMDV)-infected cattle during 2018. This study aimed to isolate and characterize the FMDVs using reverse transcription-polymerase chain reaction (RT-PCR) and gene sequencing. Materials and Methods: Forty-three FMDV-suspected clinical samples cultured on BHK-21 cell were examined, followed by virus serotype identification using RT-PCR and gene sequencing. Results: Twenty-nine (67.44%) samples were cultured on BHK-21 cell, of which 14 (32.56%) were not isolated; the 43 samples were analyzed using FMDV screening primers and serotype-specific primers. The contribution of the disease-causing serotype was serotype O of 8 (18.60%) samples, serotype A of 20 (46.51%) samples, and mixed infection (O and A) of 1 (2.33%) sample. Serotypes O and A were further characterized by phylogenetic analysis, which grouped them under East Africa 3 and Africa topotypes of genotype IV, respectively. Interestingly, serotype A was isolated for the 1st time from Keyet sub-woreda and Mulo woreda of Ethiopia, and mixed serotypes (O and A) were identified from the purchased animal. Conclusion: Molecular test result, sequencing, and phylogenetic tree reconstruction analysis revealed that the 2018 FMD outbreak in Ethiopia was caused by FMDV serotypes O and A. FMDV serotype A was the predominant strain circulating in most study areas of the country. Infections in one sample with mixed serotypes of O and A were also reported. The authors recommend a vaccine matching study of those field isolated viruses with the vaccine strain.

Ultra Rapid, One Step Molecular Detection of BCR-ABL Major and Minor Transcripts by Isothermal RT-Loop Mediated Amplification Reaction.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2515-2515
Author(s):  
Elena D'agostini ◽  
Giulia Minnucci ◽  
Giulia Amicarelli ◽  
Cinzia Pultrone ◽  
Veronica Tettamanzi ◽  
...  
Keyword(s):  
Cell Lines ◽  
Molecular Detection ◽  
Lymphoblastic Leukemia ◽  
Internal Quality ◽  
Clinical Samples ◽  
Wild Type ◽  
Rt Pcr ◽  
Automatic Instrument ◽  
Fusion Transcripts ◽  
One Step

Abstract Abstract 2515 Background: The molecular identification of the BCRABL transcripts in clinical samples is actually based on a conventional Reverse Transcription Polymerase Chain reaction (RT-PCR). Here we present a novel molecular method, based on Loop Mediated Amplification assay that, starting from RNA (RT-LAMP) in one single tube reaction ensures a rapid and simultaneous detection of either the BCR-ABL p190 or p210 fusion transcript as well as the housekeeping gene used as internal quality control. Methods: The BCR-ABL RT-LAMP is a multiplex, isothermal method for retro-transcription, amplification and detection of the Minor (p190) and Major (p210) t(9;22) transcripts and the endogenous Gusb RNA, as internal control for validation of negative results. The employment of fluorescent specific probes allows real-time monitoring of the reaction, so that the test result is obtained in a single, homogeneous step. RT-LAMP is carried out on the Liaison IAM, a 8-wells manageable instrument suitable for isothermal reactions. Liaison IAM incubates at constant temperature, monitors the fluorescent signals and the data produced can be analyzed, upon connection to up to 6 other instruments, for a throughput of 8 to 48 samples. Thanks to the three channels of fluorescence, it can monitor multiplex assays, providing elaborated final objective results with no need of further data analysis by the operator. Results: The level of sensitivity of the triplex BCR-ABL RT-LAMP has been analytically evaluated directly on serial dilutions of RNA extracted respectively from t(9;22) positive cell lines (TOM-1 for p190 or K-562 for p210) into wild type RNA from HL-60 cell line (30 replicates). The p190 and p210 transcripts have been detected and distinguished down to 10−4 and 10−5respectively within 50 minutes. The assay demonstrated 100% specificity since 70 replicates of wild type RNA from 7 cell lines resulted BCR-ABL negative and GUSb positive (internal amplification control). This assay was validated on 60 clinical samples (30 white blood cells RNA from Chronic Myeloid Leukemia, 30 mononuclear cells RNA from B-lineage Acute Lymphoblastic Leukemia). All these samples were obtained at diagnosis and were previously analyzed by conventional RT-PCR. RT-LAMP detected and identified the BCR-ABL fusion transcripts correctly in all cases with a 100% concordance with the reference method. Fully concordant results were obtained also on 30 RNA samples from patients affected by t(9;22) negative hematologic malignancies and on 30 RNA samples obtained from healthy donors in which the RT-LAMP amplified exclusively the housekeeping GUSb transcript. Conclusions: The triplex p190-p210-GUSb RT-LAMP is a one-step procedure for specific, highly sensitive and rapid molecular detection of the BCR-ABL fusion transcripts. The semi-automatic instrument Liaison IAM, simplifies the entire procedure, reduces the contamination risks deriving from the conventional, multi step RT-PCR and significantly improve the diagnostic lab routine. Disclosures: D'agostini: DIASORIN S.p.A: Employment. Minnucci:Diasorin S.p.A.: Employment. Amicarelli:DIASPRIN S.p.A.: Employment. Pultrone:DIASORIN S.p.A.: Employment. Tettamanzi:DIASORIN S.p.A.: Employment. Salmoiraghi:DIASORIN S.p.A.: Consultancy. Spinelli:DIASORIN S.p.A: Consultancy. Colotta:DIASORIN S.p.A: Employment. Rambaldi:DIASORIN S.p.A: Consultancy.

scholarly journals Seroprevalance And Molecular Detection Of Fmdv In Cattle At Savar In Bangladesh

2020 ◽  
Vol 17 (2) ◽  
pp. 67-78
Author(s):  
N Jannat ◽  
MS Rahman ◽  
E Islam ◽  
NA Rumi ◽  
M Giasuddin ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Control Measure ◽  
Clinical Signs ◽  
Effective Control ◽  
Clinical Samples ◽  
Infected Cattle ◽  
Economic Implications ◽  
Serotype A ◽  
Fmd Virus ◽  
Serotype O

The study was conducted to observe the overall seroprevalence and molecular detection of circulating FMD virus from infected cattle and efficacy of antibacterial drugs against secondary bacterial infection at Savar, Dhaka from January- December 2018. A total of 92 serum samples were collected for indirect c-ELISA to detect antibodies against non-structural proteins of FMD virus, and overall seroprevalence was 94.02%. The seroprevalence of serotype O and A was higher (95.83% and 95.83%) in male cattle than female (93.18% and 90.91%) respectively. 6 Months to 3 years aged cattle showed, significantly (p<0.01) higher seroprevalence (100%) than above 4 years age groups for serotype O (82.14%) and A (78.57%). Local cattle were more seropositive 96.88% compared to crossbred cattle 93.33% for serotype O and 91.67% for serotype A and this variation was not statistically significant (p>0.05). Among 10 clinical samples of FMD from infected cattle, 8 samples were positive for different serotypes, among them 2 each were identified as serotype A and Asia-1. On the other hand,4 samples were identified as mixed infection (1 sample of serotype O+A, 3 samples of O+Asia-1) by mRT-PCR. In this study on therapeutic intervention with sulphadimidine significantly (p<0.05) reduced the clinical signs of FMD than Gentamycin and Ampicillin. The higher seroprevalence of the disease has substantial economic implications which signify the need for devising effective control measure. However, the detection of ‘O’, Asia-1and ‘A’ serotype emphasizes the critical need for use of atrivalent vaccine in the field. SAARC J. Agri., 17(2): 67-78 (2019)

scholarly journals Molecular detection of Pasteurella multocida Type B causing haemorrhagic septicemia in cattle and buffaloes of Bangladesh

2016 ◽  
Vol 27 (2) ◽  
pp. 175-179 ◽  
Author(s):  
MS Ara ◽  
MT Rahman ◽  
M Akhtar ◽  
M Rahman ◽  
KHMNH Nazir ◽  
...  
Keyword(s):  
Morphological Study ◽  
Pasteurella Multocida ◽  
Molecular Detection ◽  
Vaccine Candidate ◽  
Clinical Samples ◽  
Effective Vaccine ◽  
Type B ◽  
Chain Reaction ◽  
Specific Primers ◽  
Polymerase Chain

Hemorrhagic septicemia (HS) is an acute septicemic disease that primarily affects cattle and buffaloes. The disease is caused by Pasteurella multocida sero types B:2 and E:2. The objective of this study was to isolate P. multocida from clinical cases and to confirm its identity using polymerase chain reaction (PCR) based approach. Clinical samples of two suspected cases of haemorrhagic septicemia of cattle and buffalo from Mymensingh and Rajshahi districts respectively were collected. Two isolates were isolated from these suspected cases and primarily identified as P. multocida based on morphological study, staining properties, and cultural and biochemical characteristics. The isolates were confirmed initially as P. multocida at genus level by PCR using genus specific primers. Later, the isolates were confirmed as P. multocida type B, the causal agent of haemorrhagic septicemia, by PCR with primers specific for P. multocida type B. These isolated organisms can be used as vaccine candidate for the production of effective vaccine against haemorrhagic septicemia.Progressive Agriculture 27 (2): 175-179, 2016

scholarly journals The Immune Correlates of Orthohantavirus Vaccine

Vaccines ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 518
Author(s):  
Joon-Yong Bae ◽  
Jin Il Kim ◽  
Mee Sook Park ◽  
Gee Eun Lee ◽  
Heedo Park ◽  
...  
Keyword(s):  
Vaccine Development ◽  
Zoonotic Transmission ◽  
Effective Vaccine ◽  
Pandemic Preparedness ◽  
Relative Paucity ◽  
Immune Correlates ◽  
Current Vaccine ◽  
Pros And Cons ◽  
Human Infections ◽  
Sense Of Urgency

Zoonotic transmission of orthohantaviruses from rodent reservoirs to humans has been the cause of severe fatalities. Human infections are reported worldwide, but vaccines have been approved only in China and Korea. Orthohantavirus vaccine development has been pursued with no sense of urgency due to the relative paucity of cases in countries outside China and Korea. However, the orthohantaviruses continuously evolve in hosts and thus the current vaccine may not work as well against some variants. Therefore, a more effective vaccine should be prepared against the orthohantaviruses. In this review, we discuss the issues caused by the orthohantavirus vaccine. Given the pros and cons of the orthohantavirus vaccine, we suggest strategies for the development of better vaccines in terms of pandemic preparedness.

scholarly journals Strategies for Vaccination: Conventional Vaccine Approaches Versus New-Generation Strategies in Combination with Adjuvants

Pharmaceutics ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 140
Author(s):  
Abdellatif Bouazzaoui ◽  
Ahmed A. H. Abdellatif ◽  
Faisal A. Al-Allaf ◽  
Neda M. Bogari ◽  
Saied Al-Dehlawi ◽  
...  
Keyword(s):  
Viral Vectors ◽  
Protein Production ◽  
Vaccine Development ◽  
Effective Vaccine ◽  
The Novel ◽  
Research Teams ◽  
Advantages And Disadvantages ◽  
Rapid Spread ◽  
New Generation ◽  
Conventional Vaccine

The current COVID-19 pandemic, caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2), has raised significant economic, social, and psychological concerns. The rapid spread of the virus, coupled with the absence of vaccines and antiviral treatments for SARS-CoV-2, has galvanized a major global endeavor to develop effective vaccines. Within a matter of just a few months of the initial outbreak, research teams worldwide, adopting a range of different strategies, embarked on a quest to develop effective vaccine that could be effectively used to suppress this virulent pathogen. In this review, we describe conventional approaches to vaccine development, including strategies employing proteins, peptides, and attenuated or inactivated pathogens in combination with adjuvants (including genetic adjuvants). We also present details of the novel strategies that were adopted by different research groups to successfully transfer recombinantly expressed antigens while using viral vectors (adenoviral and retroviral) and non-viral delivery systems, and how recently developed methods have been applied in order to produce vaccines that are based on mRNA, self-amplifying RNA (saRNA), and trans-amplifying RNA (taRNA). Moreover, we discuss the methods that are being used to enhance mRNA stability and protein production, the advantages and disadvantages of different methods, and the challenges that are encountered during the development of effective vaccines.

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