scholarly journals Detection Of Equine Arteritis Virus In The Semen Of Stallions In The Republic Of Serbia

2015 ◽  
Vol 65 (4) ◽  
pp. 557-567 ◽  
Author(s):  
Sava Lazić ◽  
Diana Lupulović ◽  
Vladimir Polaček ◽  
Miroslav Valčić ◽  
Gospava Lazić ◽  
...  
Keyword(s):  
Viral Genome ◽  
Neutralization Test ◽  
Equine Arteritis Virus ◽  
Serological Testing ◽  
Rt Pcr ◽  
High Seroprevalence ◽  
Chain Reaction ◽  
Specific Antibodies ◽  
Polymerase Chain ◽  
The Republic

AbstractThe results on serological testing of blood sera from stallions and mares used for breeding and the presence of the viral genome of Equine Arteritis Virus (EAV) in stallion semen are presented. The blood and semen samples were taken from a horse stable on the territory of the Republic of Serbia during 2012, 2013 and 2014. Detection of anti-EAV specific antibodies in blood sera was performed by the virus neutralization test (VNT), and identification of EAV genome RNA in stallion semen was done by reverse-transcriptase polymerase chain reaction (RT-PCR). In 2012, high seroprevalence of EAV was detected in the investigated stable. In total, 45% and 65 % of stallions and mares reacted positive, respectively, and the antibody titre values ranged between 2 and 10 log2. High seroprevalence was confirmed in the same animals again in 2013. Out of two stallions tested semen samples in 2013, the viral genome was detected by RT-PCR in 3 examined semen samples from a seropositive stallion, while EAV was not detected in 3 semen samples of a seronegative stallion. During 2014, 11 semen samples were collected from two seropositive stallions. Again, the presence of EAV was confirmed by RT-PCR in all 8 semen samples originating from the same stallion with the EAV genome positive semen result in 2013, whereas the virus was not detected in semen samples originating from the second anti-EAV antibody positive stallion. The presence of EAV-specific antibodies was confirmed in the blood sera of the mares inseminated with the semen of seropositive stallions in 2012 and 2013.

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 Pet rat harbouring Seoul hantavirus in Sweden, June 2013

2013 ◽  
Vol 18 (27) ◽  
Author(s):  
Å Lundkvist ◽  
J Verner-Carlsson ◽  
A Plyusnina ◽  
L Forslund ◽  
R Feinstein ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lung Tissue ◽  
Reverse Transcription ◽  
Haemorrhagic Fever ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Specific Antibodies ◽  
Polymerase Chain ◽  
Focus Reduction

We report the first detection of Seoul hantavirus (SEOV) in a pet rat in Sweden. SEOV-specific antibodies were detected in the pet rat blood by focus reduction neutralising test (FRNT), and SEOV RNA in lung tissue was confirmed by reverse transcription-nested polymerase chain reaction (RT-PCR) followed by sequencing. The discovery follows the recent reports of SEOV infected pet rats, as well as associated human cases of severe haemorrhagic fever with renal syndrome (HFRS), in England and Wales.

scholarly journals Neutralizing antibodies for SARS-CoV-2 in stray animals from Rio de Janeiro, Brazil

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0248578
Author(s):  
Helver Gonçalves Dias ◽  
Maria Eduarda Barreto Resck ◽  
Gabriela Cardoso Caldas ◽  
Alessandro Fioretti Resck ◽  
Natália Valente da Silva ◽  
...  
Keyword(s):  
Rio De Janeiro ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Serum Samples ◽  
Stray Dog ◽  
Polymerase Chain ◽  
Stray Cat ◽  
Serological Data

The epidemic of coronavirus disease 2019 (COVID-19), caused by a novel Betacoronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) became a public health emergency worldwide. Few reports indicate that owned pets from households with at least one human resident that was diagnosed with COVID-19 can be infected by SARS-CoV-2. However, the exposure to SARS-CoV-2 of pets from households with no COVID-19 cases or stray animals remains less assessed. Using real-time reverse transcriptase polymerase chain reaction (RT-PCR) and plaque reduction neutralization test (PRNT90), we investigated the infection and previous exposure of dogs and cats to SARS-CoV-2 during the ongoing COVID-19 epidemic in Rio de Janeiro, Brazil. From June to August 2020, 96 animals were sampled, including 49 cats (40 owned and 9 stray) and 47 dogs (42 owned and 5 stray). Regarding owned pets, 75.6% (62/82) belonged to households with no COVID-19 cases. Samples included serum, and rectal and oropharyngeal swabs. All swabs were negative for SARS-CoV-2 RNA, but serum samples of a stray cat and a stray dog presented neutralizing antibodies for SARS-CoV-2, with PRNT90 titer of 80 and 40, respectively. Serological data presented here suggest that not only owned pets from households with COVID19 cases, but also stray animals are being exposed to SARS-CoV-2 during the COVID-19 pandemic.

scholarly journals Circulation of bluetongue virus in goats in the Karamoja region of Uganda

2013 ◽  
Vol 84 (1) ◽  
Author(s):  
Elijah N. Mulabbi ◽  
Chrisostom Ayebazibwe ◽  
Samuel Majalija ◽  
Carrie A. Batten ◽  
Christopher A.L. Oura
Keyword(s):  
Real Time ◽  
Whole Blood ◽  
Bluetongue Virus ◽  
Rna Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
High Seroprevalence ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Karamoja Region

The presence of bluetongue virus (BTV) in indigenous goats from the Karamoja region of northern Uganda was investigated. A total of 300 goats were sampled (serum and whole blood) from five districts within the Karamoja region. The samples were analysed for the presence of bluetongue (BT) antibodies using a commercial Enzyme-linked immunosorbent assay (ELISA) and for the presence of BTV viral RNA by real-time Reverse transcription polymerase chain reaction (RT-PCR), because BTV is an RNA virus. Of the 300 goats tested, 269 (90%) were positive for BTV antibodies, indicating high levels of BTV circulation within the region. Out of the 150 whole blood samples tested for the presence of the virus by real-time RT-PCR, 84 (56%) were positive for BTV RNA. This study, which is the first of its kind in Uganda, showed a high seroprevalence of BT antibodies and active circulation of BTV in a high proportion of goats in the Karamoja region.

scholarly journals SARS-CoV-2 screening of asymptomatic women admitted for delivery must be performed with a combination of microbiological techniques: an observational study

2020 ◽  
Vol 33 (6) ◽  
pp. 415-421
Author(s):  
Maria Carmen Viñuela ◽  
Juan Antonio De León-Luis ◽  
Roberto Alonso ◽  
Pilar Catalán ◽  
Santiago Lizarraga ◽  
...  
Keyword(s):  
Clinical Care ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Systematic Screening ◽  
Specific Antibodies ◽  
Universal Testing ◽  
Microbiological Techniques ◽  
Polymerase Chain ◽  
Asymptomatic Women ◽  
Spontaneous Delivery

Introduction. The aim of this study is to assess the value of systematic screening in asymptomatic women admitted for spontaneous delivery with a combination of reverse transcription polymerase chain reaction (RT-PCR) and cycle threshold (Ct) and serum antibodies. Material and methods. Since May 6 all women admitted for spontaneous delivery underwent RT-PCR in nasopharyngeal swabs and specific antibodies IgG of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in serum that were performed as part of routine clinical care in our institution. Ct of the PCR was recorded. We analyzed the first 100 women consecutively admitted for spontaneous delivery at our institution. Results. Nine women were positive for SARS-CoV-2 in nasopharyngeal samples (9%) and 13 (13%) presented positive specific antibodies of the coronavirus. Overall, SARS-CoV-2 prior exposure was 15%. The Ct determination (RT-PCR test) of our 9 positive patients ranged from 36 to 41 cycles with a median of 40. Vaginal delivery occurred in 94% of the cases and only 6% underwent a cesarean section, always for obstetric reasons. No fetal transmission was observed and maternal and neonatal prognosis was excellent. Conclusions. During epidemic episodes in asymptomatic women in labor, universal testing with RT-PCR (considering Ct determination), and the detection of antibodies, permits a better interpretation of the results and avoid unnecessary isolation procedures.

scholarly journals Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria

2020 ◽  
Vol 13 (11) ◽  
pp. 2358-2363
Author(s):  
Ijeoma Chekwube Chukwudi ◽  
Kenneth Ikejiofor Ogbu ◽  
Pam Dachung Luka ◽  
Refiloe Petunia Malesa ◽  
Livio Edward Heath ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Tool ◽  
Viral Genome ◽  
Nasal Swab ◽  
Clinical Signs ◽  
Peste Des Petits Ruminants ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Sheep And Goats ◽  
Polymerase Chain

Background and Aim: Peste des petits ruminants (PPR) is an acute, extremely contagious transboundary viral disease of small ruminants with severe economic consequences, caused by PPR virus. Cost-effective and rapid diagnosis of the disease is essential for prompt management and control. This study aimed to compare the application of a commercial colorimetric loop-mediated isothermal amplification (cLAMP) kit and reverse transcriptase-polymerase chain reaction (RT-PCR) in the diagnosis of PPR in sheep and goats in Southeast Nigeria. Materials and Methods: Nasal swab samples were collected from West African Dwarf sheep and goats showing clinical signs suggestive of PPR (n=80) and those without any clinical signs (n=140) of the disease. The diagnosis was achieved through detection of PPR viral genome in the samples using a cLAMP kit and RT-PCR. cLAMP assay was done directly on nasal swab samples without ribosomal nucleic acid extraction. A set of six primers targeting the matrix gene protein was used for the cLAMP assay. Results: PPR viral genome was detected by both cLAMP and RT-PCR in 51 (63.8%) of the 80 samples from sheep and goats with signs suggestive of PPR while 14 (10%) of those without signs tested positive for PPR by both assay methods. There was a 100% agreement in the cLAMP and RT-PCR results. However, cLAMP was a faster, easier, and less expensive method compared to RT-PCR. Conclusion: The cLAMP assay demonstrates the potential for a point of care diagnosis in the field and a valuable diagnostic tool in areas with poor electricity supply as well as in a less equipped diagnostic laboratory. Since the reagents are affordable, cLAMP can be a diagnostic tool of choice in the detection and surveillance of PPR virus in countries with limited resources.

Investigations on the prevalence of tortoise picorna-virus in captive tortoises in Germany

2018 ◽  
Vol 46 (05) ◽  
pp. 304-308 ◽  
Author(s):  
Svenja Funcke ◽  
Michael Lierz ◽  
Susanne Paries
Keyword(s):  
Rt Pcr ◽  
Chain Reaction ◽  
Blood Samples ◽  
Specific Antibodies ◽  
Testudo Graeca ◽  
The Family ◽  
High Prevalence ◽  
Disease Pattern ◽  
Polymerase Chain ◽  
Positive Results

Summary Objective: Tortoise picornavirus (ToPV) has been speculated to play an important role in the frequently seen disease pattern of juvenile shell softening. This study aimed to determine ToPV prevalence among German tortoise collections. Material and methods: A total of 334 animals selected from 27 different collections were included. Seven species of four genera of the family Testudinidae (Testudo graeca, T. hermanni, T. marginata, T. horsfieldii, Centrochelys sulcata, Stigmochelys pardalis, Chelonoidis carbonarius) were sampled. The tortoises were clinically investigated and none of the adults showed any signs of shell softening. Seven hatchlings of a ToPV-positive T. graeca breeding pair showed retarded growth and a progressive shell weakness that resulted in death. Each animal was sampled by conjunctival, pharyngeal and cloacal swabs (990 swabs in total) and blood sampling (293 in total). All three swabs of one animal were pooled and tested by reverse transcriptase polymerase chain reaction (RT-PCR) for tortoise picornavirus RNA. Blood samples were investigated by virus neutralisation test (VNT) for specific anti ToPV antibodies. All titres equal to or higher than log2 = 2 were considered positive. Results: In total, 35 adult and 11 juvenile animals were tested positive for ToPV RNA. The serological investigation did detect specific antibodies against ToPV in 44 adult tortoises and one juvenile. In total, 76 animals were tested positive in either one of the investigations, 16 animals in both. The highest number of ToPV-positive animals was found for T. graeca, with a prevalence of 32 %. No specimens of C. carbonarius, C. sulcata, or S. pardalis were tested positive. Conclusion and clinical relevance: The results propose a predisposition in T. graeca, as well as a high prevalence of ToPV in T. graeca, whereas other species showed only single or no positive animals, but may function as virus carriers.

1504: Molecular Diagnosis and Staging of Prostate Cancer Using Clusterin in Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Molecular Diagnosis ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

Expression of Platelet Membrane Glycoprotein V in Human Megakaryocytes and Megakaryocytic Cell Lines: A Study Using a Novel Monoclonal Antibody against GPV

1994 ◽  
Vol 72 (05) ◽  
pp. 762-769 ◽  
Author(s):  
Toshiro Takafuta ◽  
Kingo Fujirmura ◽  
Hironori Kawano ◽  
Masaaki Noda ◽  
Tetsuro Fujimoto ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Immunohistochemical Analysis ◽  
Specific Protein ◽  
Platelet Membrane ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Flow Cytometric ◽  
Polymerase Chain ◽  
Megakaryocytic Cell

SummaryGlycoprotein V (GPV) is a platelet membrane protein with a molecular weight of 82 kD, and one of the leucine rich glycoproteins (LRG). By reverse transcription-polymerase chain reaction (RT-PCR), GPV cDNA was amplified from mRNA of platelets and megakaryocytic cell lines. However, since there are few reports indicating whether GPV protein is expressed in megakaryocytes as a lineage and maturation specific protein, we studied the GPV expression at the protein level by using a novel monoclonal antibody (1D9) recognizing GPV. Flow cytometric and immunohistochemical analysis indicated that GPV was detected on the surface and in the cytoplasm of only the megakaryocytes in bone marrow aspirates. In a megakaryocytic cell line UT-7, GPV antigen increased after treatment with phorbol-12-myri-state-13-acetate (PMA). These data indicate that only megakaryocytes specifically express the GPV protein among hematopoietic cells and that the expression of GPV increases with differentiation of the megakaryocyte as GPIb-IX complex.

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