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 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 The UPTAKE study: implications for the future of COVID-19 vaccination trial recruitment in UK and beyond

Trials ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Sonika Sethi ◽  
Aditi Kumar ◽  
Anandadeep Mandal ◽  
Mohammed Shaikh ◽  
Claire A. Hall ◽  
...  
Keyword(s):  
Online Survey ◽  
Age Groups ◽  
Small Towns ◽  
Vaccine Uptake ◽  
Effective Vaccine ◽  
Trial Recruitment ◽  
Trial Management ◽  
Cross Sectional ◽  
Study Results ◽  
The Future

Abstract Background Developing a safe and effective vaccine will be the principal way of controlling the COVID-19 pandemic. However, current COVID-19 vaccination trials are not adequately representing a diverse participant population in terms of age, ethnicity and comorbidities. Achieving the representative recruitment targets that are adequately powered to the study remains one of the greatest challenges in clinical trial management. To ensure accuracy and generalisability of the safety and efficacy conclusions generated by clinical trials, it is crucial to recruit patient cohorts as representative as possible of the future target population. Missing these targets can lead to reduced validity of the study results and can often slow down drug development leading to costly delays. Objective This study explores the key factors related to perceptions and participation in vaccination trials. Methods This study involved an anonymous cross-sectional online survey circulated across the UK. Statistical analysis was done in six phases. Multi-nominal logistic models examined demographic and geographic factors that may impact vaccine uptake. Results The survey had 4884 participants of which 9.44% were Black Asian Minority Ethnic (BAME). Overall, 2020 (41.4%) respondents were interested in participating in vaccine trials; 27.6% of the respondents were not interested and 31.1% were unsure. The most interested groups were male (OR = 1.29), graduates (OR = 1.28), the 40–49 and 50–59 age groups (OR = 1.88 and OR = 1.46 respectively) and those with no health issues (OR = 1.06). The least interested groups were BAME (OR = 0.43), those from villages and small towns (OR = 0.66 and 0.54 respectively) and those aged 70 and above (OR = 1.11). Conclusions In order to have a vaccination that is generalisable to the entire population, greater work needs to be done in engaging a diverse cohort of participants. Public health campaigns need to be targeted in improving trial recruitment rates for the elderly, BAME community and the less educated rural population.

scholarly journals Intention to Be Vaccinated for COVID-19 among Italian Nurses during the Pandemic

Vaccines ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 500
Author(s):  
Marco Trabucco Aurilio ◽  
Francesco Saverio Mennini ◽  
Simone Gazzillo ◽  
Laura Massini ◽  
Matteo Bolcato ◽  
...  
Keyword(s):  
Vaccine Efficacy ◽  
Online Survey ◽  
Vaccine Development ◽  
Vaccine Coverage ◽  
World Health ◽  
Vaccine Hesitancy ◽  
Effective Vaccine ◽  
Online Questionnaire ◽  
Cross Sectional ◽  
Female Sex

Background: While the COVID-19 pandemic has spread globally, health systems are overwhelmed by both direct and indirect mortality from other treatable conditions. COVID-19 vaccination was crucial to preventing and eliminating the disease, so vaccine development for COVID-19 was fast-tracked worldwide. Despite the fact that vaccination is commonly recognized as the most effective approach, according to the World Health Organization (WHO), vaccine hesitancy is a global health issue. Methods: We conducted a cross-sectional online survey of nurses in four different regions in Italy between 20 and 28 December 2020 to obtain data on the acceptance of the upcoming COVID-19 vaccination in order to plan specific interventions to increase the rate of vaccine coverage. Results: A total of 531 out of the 5000 nurses invited completed the online questionnaire. Most of the nurses enrolled in the study (73.4%) were female. Among the nurses, 91.5% intended to accept vaccination, whereas 2.3% were opposed and 6.2% were undecided. Female sex and confidence in vaccine efficacy represent the main predictors of vaccine intention among the study population using a logistic regression model, while other factors including vaccine safety concerns (side effects) were non-significant. Conclusions: Despite the availability of a safe and effective vaccine, intention to be vaccinated was suboptimal among nurses in our sample. We also found a significant number of people undecided as to whether to accept the vaccine. Contrary to expectations, concerns about the safety of the vaccine were not found to affect the acceptance rate; nurses’ perception of vaccine efficacy and female sex were the main influencing factors on attitudes toward vaccination in our sample. Since the success of the COVID-19 immunization plan depends on the uptake rate, these findings are of great interest for public health policies. Interventions aimed at increasing employee awareness of vaccination efficacy should be promoted among nurses in order to increase the number of vaccinated people.

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.

Immunological and molecular detection of rotavirus genotype in calves with gastroenteritis in Diyala-Iraq

2021 ◽  
Vol 1 (1) ◽  
pp. 001-013
Author(s):  
Ammar Talib Nasser ◽  
Abdulrazak Shafiq Hasan ◽  
Amer Khazaal Saleh ◽  
Mohammad Kassem Saleh
Keyword(s):  
Molecular Detection ◽  
Cross Sectional Study ◽  
Sectional Study ◽  
Rt Pcr ◽  
Cross Sectional ◽  
Reverse Transcription Pcr ◽  
Group A ◽  
Stool Antigen ◽  
Stool Samples ◽  
Age Range

Aim: To explore the prevalence of rotavirus infection along with the molecular detection and genotyping of group A rotavirus (RVA) among bovine calves up to 5 months old in Diyala province-Iraq. Methods: This is a cross sectional study conducted in Diyala province-Iraq during the period of 2019-2020. One hundred bovine calves with age range of 1-5 months were included in the study. All were suffering acute gastroenteritis. Serum anti-rotavirus IgM and IgG plus fecal rotavirus Ag were tested for using ELISA techniques. Stool samples positive for rotavirus Ag were submitted for reverse transcription PCR (RT-PCR) for G and P genes, followed by sequencing and genotyping thereafter. Statistical analysis was done using SPSS version 25 and P values ≤ 0.05 were considered significant. Results: The positivity rate of anti-rotavirus IgM was 80% (P = 0.0001), and that of anti-rotavirus IgG was 79% (P = 0.0001). The rotavirus stool antigen was detected in 68% of calves (P = 0.01). A total of 45 stool samples which were positive for rotavirus Ag were submitted for RT-PCR; 13 (28.9%) were positive and 32 (71.1%) were negative (P = 0.084). 10 PCR positive samples were used for sequencing and genotyping and indicated that all investigated strains belonged to G1P[8] genotype. Conclusion: The current strains analyzed belonged to the G1P[8] RVA genotypes, affirming that employment of VP7 gene polymorphism accurately yielded uniform phylogenetic distances amongst investigated rotavirus strains and that there were no noticeable assortment events between human and animal rotavirus strains in Diyala province.

scholarly journals The UPTAKE healStudy: Implications for the Future of COVID-19 Vaccination Trial Recruitment in UK and Beyond

2021 ◽  
Author(s):  
Sonika Sethi ◽  
Aditi Kumar ◽  
Anandadeep Mandal ◽  
Mohammed Shaikh ◽  
Claire Anne Hall ◽  
...  
Keyword(s):  
Online Survey ◽  
Logistic Models ◽  
Small Towns ◽  
Vaccine Uptake ◽  
Effective Vaccine ◽  
Trial Recruitment ◽  
Trial Management ◽  
Cross Sectional ◽  
Study Results ◽  
The Future

Abstract Background: Developing a safe and effective vaccine will be the principal way of controlling the COVID-19 pandemic. However, current COVID-19 vaccination trials are not adequately representing a diverse participant population in terms of age, ethnicity and comorbidities. Achieving the representative recruitment targets that are adequately powered to the study remains one of the greatest challenges in clinical trial management. To ensure accuracy and generalizability of the safety and efficacy conclusions generated by clinical trials, it is crucial to recruit patient cohorts as representative as possible of the future target population. Missing these targets can lead to reduced validity of the study results, and can often slow down drug development leading to costly delays. This study explores the key factors related to perceptions and participation in vaccination trials. Methods: This study involved an anonymous cross-sectional online survey circulated across the UK. Statistical analysis was done in six phases. Multi-nominal logistic models examined demographic and geographic factors that may impact vaccine uptake. Results: The survey had 4884 participants of which 9·44% were BAME (Black Asian Minority Ethnic). Overall 2020 (41.4%) respondents were interested in participating in vaccine trials; 27.6% of the respondents were not interested and 31.1.% were unsure. The most interested groups were male (OR=1.290), graduates (OR=1.277), the 40-49 and 50-59 age group (OR=1.880 and OR=1.460 respectively) and those with no health issues (OR=1.064). The least interested groups were BAME (OR=0.427), those from villages and small towns (OR=0.66 and 0.54 respectively), and those aged 70 and above (OR=1.11).Conclusions: In order to have a vaccination that is generalizable to the entire population, greater work needs to be done in engaging a diverse cohort of participants. Public health campaigns need to be targeted in improving trial recruitment rates for the elderly, BAME community and the less educated rural population.

scholarly journals Real-time RT-PCR diagnostics of virus causing COVID-19

2020 ◽  
Vol 13 (1) ◽  
pp. 52-63 ◽  
Author(s):  
E. V. Goncharova ◽  
A. E. Donnikov ◽  
V. V. Kadochnikova ◽  
S. A. Morozova ◽  
M. N. Boldyreva ◽  
...  
Keyword(s):  
Real Time ◽  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Medical Product ◽  
World Health ◽  
Differential Diagnostics ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Study Results ◽  
Pcr Diagnostics

Aim: the study was aimed to develop a reagent kit for the real-time RT-PCR diagnostics of virus causing COVID-19.Materials and Methods. Three target sites were chosen in the genome SARS-CoV-2. The testing included 220 samples, 48 artificially created positive samples (made from patients’ biomaterial) and 172 clinical samples (scrapes from nasal and pharyngeal cavities, bronchoalveolar lavage, expectoration, endotracheal/nasopharyngeal aspirate, feces, post-mortem material), obtained from two medical centers. Preliminary, the obtained biomaterial was analyzed with a reagent kit of comparison. The evaluation was performed with a confidential interval CI 95%. The calculation of CI for the sensitivity and specificity was made based on the distribution of χ2.Results. The authors developed a technology of novel coronavirus infection (COVID-19) real-time RT-PCR diagnostics for the application in practical healthcare and proposed the variants of testing at all the stages (preanalytical, analytical, and post-analytical, including automated results processing). The proposed reagent kit meets the requirements of the World Health Organization and the Ministry of Healthcare of the Russian Federation. The study results demonstrated high sensitivity and specificity. The sensitivity was 100% (95% CI) 95.6–100%; the specificity was 100% (95% CI) 96.7–100%.Conclusion. The proposed reagent kit was registered in the RF as a medical product; the registration certificate No. RZN 2020/9948 dated 01.04.2020. The application of the reagent kit in network laboratories will provide patients with access to testing for the virus causing COVID-19 and contribute to quick differential diagnostics, improvement of pandemic control, and accurate statistics on the spread of the virus. 

Detection of blaVIM and blaIMP Carbapenemase-Producing Pseudomonas aeruginosa from Clinical Samples in Iran

2021 ◽  
Author(s):  
Mohammad Reza Zolfaghari
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cross Sectional Study ◽  
Clinical Samples ◽  
Sectional Study ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Cross Sectional ◽  
High Prevalence ◽  
Polymerase Chain ◽  
High Level

Background and Aims: The prevalence of carbapenemase-producing Pseudomonas aeruginosa (P. aeruginosa) strains has been recently reported worldwide. Therefore, accurate and rapid detection of carbapenemase-producing isolates is essential. So, this study aimed to detect blaVIM and blaIMP carbapenemase-producing strains using the modified Hodge test (MHT) and reverse transcription-polymerase chain reaction (RT-PCR). Materials and Methods: In this cross-sectional study, P. aeruginosa  strains were collected from clinical samples (blood, urine, wound, and other liquids body) in Firoozgar and Shahid Motahari Hospitals in Tehran and Velayat Hospital in Rasht Province, from May to December 2018. After identifying the isolates using the standard microbial tests, carbapenemase-producing strains were isolated by the modified hodge test. After that, the detection of blaVIM and blaIMP genes was performed by RT-PCR technique. Results: One hundred P. aeruginosa were isolated from different clinical samples. Among these, 74 (74%) isolates were considered as carbapenemase positive using MHT. The frequencies of blaVIM and blaIMP genes were obtained as 83% and 11%, respectively. Conclusions: The results of this study indicate a high level of resistance to most of the antibiotics tested and a high prevalence of blaVIM gene in P. aeruginosa strains.

Reverse Transcriptase PCR (Rt-PCR) for Detection of Dengue and Chikungunya Virus of Mosquito Aedes aegypti in Sokaraja

2020 ◽  
Vol 2 (1) ◽  
pp. 56
Author(s):  
Dyah Retno Annisa ◽  
Endang Srimurni Kusmintarsih ◽  
Trisnowati Budi Ambarningrum
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Chikungunya Virus ◽  
Sampling Method ◽  
Molecular Detection ◽  
Transmission Risk ◽  
Virus Infections ◽  
Rt Pcr ◽  
Cross Sectional

dengue fever (DF). Meanwhile, chikungunya virus causes Chikungunya fever (CF). These diseases involve three organisms, namely virus, mosquito Aedes sp., and human. The transmission of dengue and chikungunya virus is related to the population of Ae. aegypti. Banyumas regency is one of the regions with many cases of dengue and chikungnya virus infections, particularly in Purwokerto, Sokaraja, and Cilongok sub-district. Up to this time, there is no medicine and vaccine provided to treat these viruses effectively. Thus, detection of virus inside vector will be effectively performed in order to predict the transmission risk of dengue and chikungunya virus. This research aimed to know the molecular detection of dengue and chikungunya virus on adult Ae.aegypti mosquito in Sokaraja Region, Banyumas Regency. Survey was done by a cross-sectional method in Sokaraja sub-district from May 2019 – March 2019. Furthermore, technical sampling that used was purposive sampling method of adult Ae.aegypti using BG-Sentital Trap, followed by molecular detection of dengue virus using Two-step RT-PCR and chikungunya gene virus using RT-PCR. Molecular detection of DENV and CHIKV of mosquitoes which collected from Sokaraja region showed negative result

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