scholarly journals Identification et caractérisation des isolats d'orbivirus des îles de la Réunion et de la Martinique

2009 ◽  
Vol 62 (2-4) ◽  
pp. 149
Author(s):  
D. Vitour ◽  
Corinne Sailleau ◽  
Emmanuel Breard ◽  
Stéphan Zientara
Keyword(s):  
Sequence Analysis ◽  
Clinical Symptoms ◽  
Enzyme Linked Immunosorbent Assay ◽  
La Réunion ◽  
Rt Pcr ◽  
Polymerase Chain ◽  
The One ◽  
Almost All ◽  
Haemorrhagic Disease

At the beginning of 2009, bluetongue (BT)-like clinical symptoms were reported in cattle on the French island of La Réunion (Indian Ocean). One hundred and twenty-three cows were blood tested for the presence of BT and/or epizootic haemorrhagic disease virus (EHDV) ribonucleic acid (RNA) by group specific reverse transcriptase polymerase chain reaction (RT-PCR). EHDV RNA was detected in 111 samples (90.2%), among which five were also positive for BTV RNA. Sequence analysis of EHDV segment 7 revealed that this circulating strain seemed to be similar to the one isolated in 2003 (99.8% nucleotide identity). The determination of the nucleotide sequence of segment 2 is under investigation. The vironeutralization test (VNT), serotype-specific RT-PCR, as well as sequence analysis identified the isolated BTV strain as serotype 2. These data showed that an EHDV outbreak occurred over the last winter in La Réunion, and it was concomitant to circulation of BTV. Epidemic or enzootic features of both these viruses are not yet known. Since this outbreak, molecular and serological tools specific to EHDV have been or are being developed. Three years ago, 30 healthy head of cattle moved from Metropolitan France to the French Martinique Island (Caribbean Basin) and were distributed in four different farms. Animals were sampled (blood and serum) every 10 days until day 30 and tested for BTV infection by the enzyme-linked immunosorbent assay (ELISA) or RT-PCR assays. Unexpectedly, almost all animals became BTV positive within 20 days. Whenever possible, virus isolation on eggs and baby hamster kidney (BHK) cell cultures were performed. Interestingly, seven BT strains belonging to seven distinct serotypes (BTV-2, 9, 10, 17, 18, 22, 24) were isolated. The coding sequence of segments 7, 8, 9 and 10 of these seven serotypes was obtained, as well as a portion of segment 2. The phylogenetic analysis revealed an unprecedented divergence of these strains with other known BTV sequences.

scholarly journals Isolation, characterization and sequence determination of Noda virus from sea bass (Dicentrarchus labraxL.) reared in freshwater and marine facilities in Greece

2017 ◽  
Vol 56 (2) ◽  
pp. 105
Author(s):  
E. XYLOURI (Ε. ΞΥΛΟΥΡΗ) ◽  
J. KOTZAMANIS (Ι. ΚΟΤΖΑΜΑΝΗΣ) ◽  
F. ATHANASSOPOULOU (ΑΘΑΝΑΣΟΠΟΥΛΟΥ Φ.) ◽  
L. DONG ◽  
A. ARGYROKASTRITIS (ΑΛ. ΑΡΓΥΡΟΚΑΣΤΡΙΤΗΣ) ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Dicentrarchus Labrax ◽  
Sea Bass ◽  
Rt Pcr ◽  
Sequence Determination ◽  
Tissue Samples ◽  
Diseased Fish ◽  
Polymerase Chain ◽  
Virus Isolates

The present study aims το characterize Noda virus isolates from freshwater and marine fish, specifically from species Dicentrarchus labrax (sea bass). Previous works reported the detection of Noda virus only by using PCR, in cultured Dicentrarchus labrax showing clinical symptoms. Here we describe the isolation of Noda virus from symptomatic sea bass cultured in sea and freshwater facilities in Greece. The virus was isolated in the continuous cell line SSN -1, exhibiting characteristic cytopathic effect, vacuolation and degeneration of the monolayer. In parallel, in all cerebral tissue samples isolated from infected individuals, a 255 bp viral fragment has been detected, using Reverse Transcription - Polymerase Chain Reaction (RT-PCR) and nested PCR. Comparison of the amplified sequence was detected in diseased fish in European farms and in other piscine species revealed a high nucleotide homology.

scholarly journals Development of a reverse transcription polymerase chain reaction for the detection of severe fever with thrombocytopenia syndrome virus from suspected infected animals

2020 ◽  
Author(s):  
Eun-sil Park ◽  
Osamu Fujita ◽  
Masanobu Kimura ◽  
Akitoyo Hotta ◽  
Koichi Imaoka ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Neutralizing Antibodies ◽  
Clinical Symptoms ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Severe Fever

AbstractBackgroundSevere fever with thrombocytopenia syndrome virus (SFTSV) causes severe hemorrhagic fever in humans and cats. Clinical symptoms of SFTS-infected cats resemble to those of SFTS patients and SFTS-contracted cats shows high levels of viral RNA loads in the serum and body fluids. Due to the risk of direct infection from SFTS-infected cats to human, it is important to diagnose SFTS-suspected animals.Methodology/Principle findingsFour primer sets were newly designed from consensus sequences constructed by 108 strains of SFTSV. A reverse transcription polymerase chain reaction (RT-PCR) with these four primer sets were successfully and specifically detected several clades of SFTSV. Their limits of detection are 1-10 copies/reaction. By this RT-PCR, 5 cat cases among 56 SFTS-suspected animal cases were diagnosed as SFTS. From these cats, IgM or IgG against SFTSV were detected by enzyme-linked immunosorbent assay (ELISA), but not neutralizing antibodies by plaque reduction neutralization titer (PRNT) test. This phenomenon is similar to those of fatal SFTS patients.Conclusion/SignificanceThis newly developed RT-PCR could detect SFTSV RNA of several clades from SFTS-suspected animals. In addition to ELISA and PRNT test, the useful laboratory diagnosis systems of SFTS-suspected animals has been made in this study.Author summaryThis study developed RT-PCR to detect SFTS animal cases. This assay could detect SFTSV RNA belonging to different clades. Cats diagnosed as SFTS had IgM or IgG, but not neutralizing antibodies. SFTS cat cases were distributed in the area where SFTS patients have been reported highly, indicating the establishment of the circulation of SFTSV in the environment. These diagnostic assays could be helpful tools to detect and not to miss SFTS animal cases.

scholarly journals Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Genome ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Kappa Value ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Level Of Agreement

<b><i>Background:</i></b> Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. <b><i>Methods:</i></b> A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. <b><i>Results:</i></b> S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (<i>p</i> = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. <b><i>Conclusions:</i></b> The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

scholarly journals Monitoring Current-Season Spread of Potato virus Y in Potato Fields Using ELISA and Real-Time RT-PCR

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 641-644 ◽  
Author(s):  
Manphool S. Fageria ◽  
Mathuresh Singh ◽  
Upeksha Nanayakkara ◽  
Yvan Pelletier ◽  
Xianzhou Nie ◽  
...  
Keyword(s):  
Real Time ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Current Season ◽  
Seed Market ◽  
Polymerase Chain ◽  
Time Of Harvest

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.

scholarly journals Epidemiology and Control of Congo Fever in Sacrificial Animals of Pakistan

2019 ◽  
Vol 1 (2) ◽  
Author(s):  
Hafiz Muhammad Rizwan ◽  
Muhammad Sohail Sajid ◽  
Haider Abbas ◽  
Muhammad Fiaz Qamar ◽  
Qaiser Akram
Keyword(s):  
Tick Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Many Body ◽  
Rt Pcr ◽  
Body Tissues ◽  
Crimean Congo Haemorrhagic Fever ◽  
Polymerase Chain ◽  
And Control ◽  
Successful Prevention ◽  
Venereal Transmission

The cases and deaths due to Crimean-Congo haemorrhagic fever (CCHF) [49] virus commonly known as Congo virus (fatality rate 15%) have been reported throughout Pakistan from the last five years especially during religious occasion, Eid-ul-Azha. The annual increase in death rates due to CCHF demonstrate the importance of awareness of Congo fever at academia as well as public level. The symptoms of Congo fever which appear one to nine days after tick bite, include sudden high fever, muscle aches, abdominal pain, headache, dizziness, sore eyes, jaundice, mood swings, confusion, aggression, and sensitivity to light. The other signs include sore throat, joint pain, vomiting, diarrhea, hemorrhages, and bleeding from skin and large intestine. The Infection has been reported in many species of wild as well as domestic animals including hares, cattle, sheep, goats, dogs, mice and hedgehogs. At least 31 species of Hyalomma, Boophilus, Rhipicephalus, Dermacentor (Ixodidae: hard ticks) act as vector of CCHF in which transovarial, transstadial and venereal transmission occurs. The virus attacks the immune system of the host and influences the immune cells. The Congo fever virus can be isolated from blood, plasma and many body tissues (kidneys, liver, spleen, lungs, brain and bone marrow). Mice inoculation, enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR) can be used for detection of the infection. Furthermore, IgM and IgG antibodies against CCHFV can also be detected and quantified. Education of general public, tick control with acaricides, use of anti-CCHFV immunoglobulin, usage of approved repellents to prevent tick bites, wearing neutral-coloured garments, application of a permethrin spray to the clothing, avoiding tall grasses and shrubs, applying sunscreen, avoiding direct contact with the blood or tissues of animals are the factors for successful prevention of the infection.

scholarly journals Detection of Cherry leaf roll virus and Strawberry latent ring spot virus by one-step RT-PCR

2009 ◽  
Vol 45 (No. 4) ◽  
pp. 140-143 ◽  
Author(s):  
S. Kumari
Keyword(s):  
Leaf Roll ◽  
Rt Pcr ◽  
Spot Virus ◽  
Cherry Leaf Roll Virus ◽  
One Step ◽  
Polymerase Chain ◽  
The One ◽  
Ring Spot ◽  
Screen House ◽  
Xiphinema Diversicaudatum

A one-step reverse transcription-polymerase chain reaction (RT-PCR) protocol was developed and used for the detection of <i>Cherry leaf roll virus</i> (CLRV) and <i>Strawberry latent ring spot virus</i> (SLRSV). The protocol was used to test infected screen house plants and also plants from orchards and vineyards where the vector (<i>Xiphinema diversicaudatum</i>) of SLRSV was detected from the soil. The one-step RT-PCR protocol is rapid and sensitive and has the potential to be used for the diagnosis of CLRV and SLRSV in routine diagnostic laboratories.

scholarly journals Detection of Border Disease Virus in Fetuses, Stillbirths, and Newborn Lambs from Natural and Experimental Infections

2009 ◽  
Vol 21 (3) ◽  
pp. 331-337 ◽  
Author(s):  
Ana L. García-Pérez ◽  
Esmeralda Minguijón ◽  
Jesús F. Barandika ◽  
Gorka Aduriz ◽  
Inés Povedano ◽  
...  
Keyword(s):  
Disease Virus ◽  
Experimental Infection ◽  
Clinical Symptoms ◽  
Pcr Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Border Disease Virus ◽  
Rt Pcr ◽  
Genotype 4 ◽  
Antigen Elisa ◽  
Border Disease

The purpose of the present study was to evaluate the use of enzyme-linked immunosorbent assay (ELISA) antigen detection in blood or fetal fluids and reverse transcription polymerase chain reaction (RT-PCR) amplification in tissues for routine laboratory diagnosis of Border disease virus (BDV) infection. Samples from 67 fetuses, 6 stillbirths, and 11 lambs from 25 commercial flocks with suspicion of BDV abortion and 3 fetuses, 7 stillbirths, and 15 lambs obtained from an experimental infection with a local isolate (BDV genotype 4) were investigated. Presence of BDV was detected by RT-PCR in 7.9% of fetuses, 50% of stillbirths, and 50% of lambs from the commercial flocks analyzed, corresponding to 8 of the 25 farms (32%). A similar percentage of the lambs and stillbirths from the experimental infection were positive by RT-PCR of tissue samples (54.5%), and the highest positivity was detected in lymph node, thyroid gland, and kidney. The current study revealed that RT-PCR analysis of stillbirths and lambs with clinical symptoms is more suitable than the analysis of fetuses to confirm the presence of BDV in a flock. Pestiviral antigen was detected by antigen ELISA in a high proportion of fetuses (24/58) and stillbirths (3/4) from commercial flocks, but in lambs, the presence of colostral antibodies masked the detection of the antigen by ELISA. Nevertheless, in lambs from the experimental infection that were not fed colostrum, antigen ELISA was less efficient than RT-PCR in detecting viral presence in stillbirths and lambs. Antigen ELISA is therefore recommended for fetuses with advanced autolysis that can adversely affect RNA integrity.

scholarly journals Serological and Molecular Detection of Hepatitis B virus among patients referred to Kurdistan Center for Hepatology and Gastroenterology in Sulaimani City/Kurdistan Region of Iraq

2020 ◽  
Vol 4 (2) ◽  
pp. 117
Author(s):  
Raz Sirwan Abdulla ◽  
Salih Ahmed Hama
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hbv Dna ◽  
Molecular Techniques ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
B Virus ◽  
Polymerase Chain ◽  
Pcr Techniques ◽  
Positive Results

Hepatitis B virus infection is caused by the hepatitis B virus, a major global health problem. This infection can lead to chronic conditions, followed by cirrhosis and hepatocellular carcinoma (HCC). The current study was aimed to detect HBV using serological and molecular techniques. During 2019, 300 blood samples were collected from Kurdistan Center for Hepatology and Gastroenterology in Sulaimani city. Enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) techniques were used for the detection of HBsAg and HBV DNA, respectively. Obtained results were revealed that 92 out of 300 tested patients (30.66%) seropositive for HBsAg. Among 92 seropositive patients, 53 were shown positive results for HBV DNA by RT-PCR. Dental clinic visiting and dialysis were among the important risk factors for HBV transmission. The vast majority of positive results were among males. Smokers showed relatively high rates of positive results. One-third of the referred patients who had liver complaints were positive for HBsAg. More than half of the seropositive patients showed RT-PCR positive results. It was concluded that the molecular method (RT-PCR) is more sensitive and gives a more accurate result than serology (ELISA). Therefore, it can be used as a diagnostic tool for HBV detection.

scholarly journals Modern biomarker cardiotrophin-1 in the diagnosis of myocardial diastolic function in men with essential hypertension

2020 ◽  
Vol 24 (3) ◽  
pp. 460-464
Author(s):  
M. O. Matokhniuk ◽  
V. M. Zhebel ◽  
L. V. Kulchevich ◽  
O. K. Shevchuk
Keyword(s):  
Heart Failure ◽  
Essential Hypertension ◽  
Chronic Heart Failure ◽  
Diastolic Function ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Standard Statistical Package ◽  
Polymerase Chain ◽  
Mathematical Processing

Annotation. The aim of the study was to improve the diagnosis of myocardial diastolic function in men with essential hypertension and chronic heart failure with different variants of the cardiotrophin-1 gene. 100 people aged 40–60 were examined: 50 men with asymptomatic essential hypertension and 50 men with essential hypertension complicated by chronic heart failure. Genotyping of the cardiotrophin-1 gene was performed by polymerase chain reaction. Determination of the concentration of cardiotrophin-1 was performed by enzyme-linked immunosorbent assay. Structural and functional parameters of the myocardium were evaluated using ultrasound of the heart. Mathematical processing of the material was performed on a personal computer using the standard statistical package SPSS 10. It was found that in persons with essential hypertension and chronic heart failure, carriers of the GA + AA variant of the cardiotrophin-1 gene (rs8046707) have significantly higher plasma peptide levels with diastolic dysfunction (p <0.05).

scholarly journals Point-of-Care Diagnostic Tools for Surveillance of SARS-CoV-2 Infections

2021 ◽  
Vol 9 ◽  
Author(s):  
Dhanasekaran Sakthivel ◽  
David Delgado-Diaz ◽  
Laura McArthur ◽  
William Hopper ◽  
Jack S. Richards ◽  
...  
Keyword(s):  
Large Scale ◽  
Clinical Symptoms ◽  
Point Of Care ◽  
Cost Effective ◽  
Nucleic Acid Amplification ◽  
Diagnostic Tools ◽  
Rt Pcr ◽  
Serological Tests ◽  
Polymerase Chain ◽  
Effective Public Health

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a recently emerged and highly contagious virus that causes coronavirus disease 2019 (COVID-19). As of August 24, 2021, there were more than 212 million confirmed COVID-19 cases and nearly 4.4 million deaths reported globally. Early diagnosis and isolation of infected individuals remains one of the most effective public health interventions to control SARS-CoV-2 spread and for effective clinical management of COVID-19 cases. Currently, SARS-CoV-2 infection is diagnosed presumptively based on clinical symptoms and confirmed by detecting the viral RNA in respiratory samples using reverse transcription polymerase chain reaction (RT-PCR). Standard RT-PCR protocols are time consuming, expensive, and technically demanding, which makes them a poor choice for large scale and point-of-care screening in resource-poor settings. Recently developed isothermal nucleic acid amplification tests (iNAAT), antigen and/or serological tests are cost-effective to scale COVID-19 testing at the point-of-care (PoC) and for surveillance activities. This review discusses the development of rapid PoC molecular tools for the detection and surveillance of SARS-CoV-2 infections.

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