Investigation of Rotavirus infection in sheep usingserological and molecular techniques

2016 ◽  
Author(s):  
Volkan Yilmaz. ◽  
M.Ozkan Timurkan ◽  
Nuvit Coskun ◽  
Yakup Yildirim
Keyword(s):  
Molecular Detection ◽  
Molecular Techniques ◽  
Enzyme Linked Immunosorbent Assay ◽  
Small Scale ◽  
Family Farms ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Rotavirus Infection ◽  
Scale Family ◽  
Polymerase Chain

In this study, serological and molecular research was conducted on the Rotavirus infection in domestic breeds of sheep at 2–3 years of age. The sheep included in the study were raised on small scale family units of less than 20 sheep per unit, in central Kars province and its districts (Susuz, Arpaçay, Kagizman and Selim) in the Northeast Anatolia region of Turkey. The blood and fecal samples were collected randomly from 450 sheep. They were analyzed for the presence of Rotavirus and the antibody against the virus using enzyme-linked immunosorbent assay (ELISA). The highest seropositive ratio (73.46%) was found in central Kars province. The seroprevalence of Rotavirus in sheep raised in the Kars region was determined to be 55.33%. Rotavirus was not detected in fecal samples with ELISA. Molecular detection of Rotavirus from fecal samples was done by reverse transcription polymerase chain reaction (RT-PCR) technique using specific generic primers for VP6 protein. Rotavirus could not be detected in RT-PCR. The data that were obtained showed that the infection spreads on small scale family farms. Based on this information, recommendations were made for controlling Rotavirus infection.

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 Seeing beyond a Dilated Proventriculus: Diagnostic Tools for Proventricular Dilatation Disease in Psittacine Birds

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3558
Author(s):  
Jeann Leal de Araújo ◽  
Raquel Rubia Rech
Keyword(s):  
Molecular Techniques ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Postmortem Diagnosis ◽  
Concise Review ◽  
Life Threatening ◽  
Polymerase Chain ◽  
Proventricular Dilatation Disease ◽  
Diagnostic Outcomes

Proventricular dilatation disease (PDD) is a life-threatening neurological disease caused by parrot bornaviruses (PaBVs) that affects several species worldwide. PDD can be clinically manifested as either a central nervous system condition or a gastrointestinal condition if the nerves and ganglia of the gastrointestinal tract are compromised. We intend to provide a concise review for veterinary clinicians and diagnosticians with focus on the main tools available for PDD diagnosis, including gross and histopathology, immunohistochemistry, molecular techniques and serology. We suggest that a combination of different strategies can increase the success of diagnostic outcomes, as tools such as reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) can be implemented for identification of bornaviral infections in live patients, and gross pathology, histopathology, immunohistochemistry and RT-PCR can provide reliable results for postmortem diagnosis of PDD.

scholarly journals Isolation and Characterization of Coronavirus and Rotavirus Associated With Calves, in Central Part of Oromia, Ethiopia

2020 ◽  
Author(s):  
Umer Seid Geletu ◽  
Fufa Dawo Bari ◽  
Munera Ahmednur Usmael ◽  
Asamino Tesfaye
Keyword(s):  
Diarrheal Disease ◽  
Addis Ababa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infection Prevalence ◽  
Rt Pcr ◽  
Cross Sectional ◽  
Fecal Samples ◽  
Rotavirus Infection ◽  
Isolation And Characterization

Abstract Background: Coronavirus and Rotavirus are most commonly associated etiologies for calves’ diarrhea resulting in loss of productivity and economy of farmers. However, various facets of diarrheal disease caused by coronavirus and rotavirus in calves in Ethiopia are inadequately understood. A cross sectional study was conducted with the aim of isolation and molecular characterization of coronavirus and rotavirus from calves in central part of Oromia (Bishoftu, Sebata, Holeta and Addis Ababa), Ethiopia from November 2018 to May 2019. The four study areas were purposively selected and fecal samples were collected by simple random sampling for diagnosis of coronavirus and rotavirus infection by using antigen detection Enzyme linked immunosorbent assay (Ag-ELISA) kit. In addition, this study was carried out to have insight in prevalence and associated risk factors of coronavirus and rotavirus infection in calves. Result: During the study 83 diarrheic and 162 non-diarrheic fecal samples collected from calves less than 4 weeks of age were screened for coronavirus and rotavirus. Of the 83 diarrheic samples, 1 sample (1.2%) was positive for coronavirus antigen (Ag) and 6 samples (7.2%) were found to be positive for rotavirus antigen (Ag) by Ag-ELISA. All the non-diarrheic samples were negative for both coronavirus and rotavirus Ag. The overall prevalence of coronavirus and rotavirus infection in calves were estimated as 0.4% (1/245) and 2.45% (6/245) respectively. All samples (7) of ELISA test positive of both coronavirus and rotavirus were propagated in Madin Darby bovine kidney cells (MDBK). After 3 subsequent passages, progressive cytopathic effect (CPE) i.e. rounding, detachment as well as destruction of mono-layer cell of five sample (1 sample of coronavirus and 4 sample of rotavirus) (71.4%) were observed. At the molecular stage, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to determine the presence of coronavirus and rotavirus nucleic acid by using specific primers. The 5 samples that were coronavirus and rotavirus antigen positive by ELISA and develop CPE on cell culture were also positive on RT-PCR technique. Infection prevalence peaked have been obtained at 1st and 2nd weeks of age in male calves. Conclusion: Diarrheal disease caused by coronavirus and rotavirus has a great health problem in calves that interrupts production benefits with reduced weight gain and increased mortality, and its potential for zoonotic spread. So the present findings show coronavirus and rotavirus infection in calves in Ethiopia that needs to be addressed by practicing early colostrums feeding in newborn calves, using vaccine, or improving livestock management.

Isolation Coronavirus and Rotavirus Associated With Calves, in Central Part of Oromia, Ethiopia

2020 ◽  
Author(s):  
Umer Seid ◽  
Fufa Dawo ◽  
Munera Ahmednur ◽  
Asamino Tesfaye
Keyword(s):  
Diarrheal Disease ◽  
Addis Ababa ◽  
Simple Random Sampling ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infection Prevalence ◽  
Rt Pcr ◽  
Cross Sectional ◽  
Fecal Samples ◽  
Rotavirus Infection

Abstract Background: Coronavirus and Rotavirus are most commonly associated etiologies for calves’ diarrhea resulting in loss of productivity and economy of farmers. However, various facets of diarrheal disease caused by coronavirus and rotavirus in calves in Ethiopia are inadequately understood. A cross sectional study was conducted with the aim of isolation and molecular characterization of coronavirus and rotavirus from calves in central part of Oromia (Bishoftu, Sebata, Holeta and Addis Ababa), Ethiopia from November 2018 to May 2019. The four study areas were purposively selected and fecal samples were collected by simple random sampling for diagnosis of coronavirus and rotavirus infection by using antigen detection Enzyme linked immunosorbent assay (Ag-ELISA) kit. In addition, this study was carried out to have insight in prevalence and associated risk factors of coronavirus and rotavirus infection in calves. Result: During the study 83 diarrheic and 162 non-diarrheic fecal samples collected from calves less than 4 weeks of age were screened for coronavirus and rotavirus. Of the 83 diarrheic samples, 1 sample (1.2%) was positive for coronavirus antigen (Ag) and 6 samples (7.2%) were found to be positive for rotavirus antigen (Ag) by Ag-ELISA. All the non-diarrheic samples were negative for both coronavirus and rotavirus Ag. The overall prevalence of coronavirus and rotavirus infection in calves were estimated as 0.4% (1/245) and 2.45% (6/245) respectively. All samples (7) of ELISA test positive of both coronavirus and rotavirus were propagated in Madin Darby bovine kidney cells (MDBK). After 3 subsequent passages, progressive cytopathic effect (CPE) i.e. rounding, detachment as well as destruction of mono-layer cell of five sample (1 sample of coronavirus and 4 sample of rotavirus) (71.4%) were observed. At the molecular stage, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to determine the presence of coronavirus and rotavirus nucleic acid by using specific primers. The 5 samples that were coronavirus and rotavirus antigen positive by ELISA and develop CPE on cell culture were also positive on RT-PCR technique. Infection prevalence peaked have been obtained at 1st and 2nd weeks of age in male calves. Conclusion: Diarrheal disease caused by coronavirus and rotavirus has a great health problem in calves that interrupts production benefits with reduced weight gain and increased mortality, and its potential for zoonotic spread. So the present findings show coronavirus and rotavirus infection in calves in Ethiopia that needs to be addressed by practicing early colostrums feeding in newborn calves, using vaccine, or improving livestock management.

scholarly journals Performance of Latex agglutination, ELISA and RT-PCR for diagnosis of Rotavirus infection

2018 ◽  
Vol 90 (2) ◽  
Author(s):  
Hosna Hamzavi ◽  
Azarakhsh Azaran ◽  
Manoochehr Makvandi ◽  
Sahar Karami ◽  
Mohammad Roayaei Ardakani ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Acute Gastroenteritis ◽  
Laboratory Diagnosis ◽  
Latex Agglutination ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Rotavirus Infection ◽  
Polymerase Chain ◽  
Major Factors ◽  
Positive Results

The rotavirus is one of the major factors of inducing the acute gastroenteritis infection in children under 5 years of age. The laboratory diagnosis is progress and bringing it under control as well as avoiding its diffusion. The purpose of the present study was to determine the performance of enzyme linked immunosorbent assay (ELISA) and Latex agglutination (LA) tests against reverse transcription-polymerase chain reaction (RT-PCR) for evaluating the children’s acute gastroenteritis by rotavirus. One hundred feces specimens were collected from February to May 2014 and analyzed by LA, ELISA and RT-PCR. In this study, the positive results for rotavirus detected by ELISA, LA and RT-PCR were 37, 43 and 27%, respectively. In addition, the result showed that the sensitivity and specificity of ELISA and LA were 74 and 85%, respectively, when compared to RT-PCR. For laboratory detection of Rotavirus infection, RT-PCR has the highest sensitivity and specificity but because of the high costs, ELISA and LA based kits with good performance, as shown by this study, can be preferred for the routine use.

scholarly journals Detection of Bovine Torovirus and other Enteric Pathogens in Feces from Diarrhea Cases in Cattle

2003 ◽  
Vol 15 (3) ◽  
pp. 205-212 ◽  
Author(s):  
Armando E. Hoet ◽  
Paul R. Nielsen ◽  
Mustafa Hasoksuz ◽  
Christopher Thomas ◽  
Thomas E. Wittum ◽  
...  
Keyword(s):  
Etiologic Agent ◽  
Enteric Pathogens ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Salmonella Spp ◽  
Diagnostic Laboratory ◽  
Fecal Samples ◽  
Polymerase Chain

The objectives of this study were to determine the prevalence of bovine torovirus (BoTV) in bovine fecal samples from diarrhea cases submitted to the Ohio Animal Disease Diagnostic Laboratory (ADDL) and to assess if a relationship exists between BoTV and the other enteric pathogens detected. From November 1999 to May 2001, 259 specimens from 53 calves (≤6 months old), 27 young adults (≤2 years), 125 adults (≥2 years), and 54 animals of unknown age were examined by an antigen-capture enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase-polymerase chain reaction (RT-PCR) assay developed to detect BoTV Testing for other enteric pathogens was performed by ADDL, and the results were analyzed with the BoTV data. The BoTV was detected using ELISA or RT-PCR in 9.7% (25/259) of the clinical samples, 56% (14/25) of which were from calves ( P < 0.001) representing 26.4% (14/53) of the calves tested. Of the BoTV-positive calves, 71% (10/14) were less than 3 weeks of age. In 11/25 positive specimens, BoTV was the only pathogen detected among those examined. Other enteric organisms detected alone or in combination with BoTV in calf samples were rotavirus, coronavirus, Salmonella spp., Cryptosporidium spp., and Giardia spp.; but no consistent association between BoTV and these organisms was observed. In summary, BoTV was detected in fecal samples from cattle with diarrhea, principally in young calves less than 3 weeks of age. Future studies of infectious diarrhea in cattle should also include assays for this etiologic agent.

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 Prevalence and molecular detection of shiga toxin producing Escherichia coli from diarrheic cattle

2016 ◽  
Vol 14 (1) ◽  
pp. 63-68 ◽  
Author(s):  
MM Akter ◽  
S Majumder ◽  
KH MNH Nazir ◽  
M Rahman
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Molecular Detection ◽  
Chain Reaction ◽  
Specific Primers ◽  
Fecal Samples ◽  
E Coli ◽  
Uremic Syndrome ◽  
Polymerase Chain ◽  
Positive Isolate

Shiga toxin-producing Escherichia coli (STEC) are zoonotically important pathogen which causes hemorrhagic colitis, diarrhea, and hemolytic uremic syndrome in animals and humans. The present study was designed to isolate and identify the STEC from fecal samples of diarrheic cattle. A total of 35 diarrheic fecal samples were collected from Bangladesh Agricultural University (BAU) Veterinary Teaching Hospital. The samples were primarily examined for the detection of E. coli by cultural, morphological and biochemical characteristics, followed by confirmation of the isolates by Polymerase Chain Reaction (PCR) using gene specific primers. Later, the STEC were identified among the isolated E. coli through detection of Stx-1 and Stx-2 genes using duplex PCR. Out of 35 samples, 25 (71.43%) isolates were confirmed to be associated with E. coli, of which only 7 (28%) isolates were shiga toxin producers, and all of them were positive for Stx-1. However, no Stx-2 positive isolate could be detected. From this study, it may be concluded that cattle can act as a reservoir of STEC which may transmit to human or other animals.J. Bangladesh Agril. Univ. 14(1): 63-68, June 2016

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.

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