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

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.

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Evaluation of the Reverse Transcription-Polymerase Chain Reaction/Probe Test of Serum Samples and Immunohistochemistry of Skin Sections for Detection of Acute Bovine Viral Diarrhea Infections

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870201400405 ◽

2002 ◽

Vol 14 (4) ◽

pp. 303-307 ◽

Cited By ~ 27

Author(s):

Julia F. Ridpath ◽

Sharon K. Hietala ◽

Steve Sorden ◽

John D. Neill

Keyword(s):

Polymerase Chain Reaction ◽

Polymerase Chain Reaction Amplification ◽

Clinical Signs ◽

Bovine Viral Diarrhea ◽

Rt Pcr ◽

Chain Reaction ◽

Serum Samples ◽

Viral Diarrhea ◽

Persistent Infections ◽

Polymerase Chain

Bovine viral diarrhea viruses (BVDV) cause both acute and persistent infections. While diagnostic tests have been designed to detect animals persistently infected (PI) with BVDV, the reliability of these tests in detecting acute BVDV infections is not known. It is also possible that acute BVDV infections may be confused with persistent infections in surveys for PI animals. In this study, 2 tests presently in use in diagnostic laboratories to test for PI animals, polymerase chain reaction amplification followed by probe hybridization (RT-PCR/probe) of serum samples and immunohistochemical detection of viral antigen in skin biopsies (IHC), were evaluated for their ability to detect acute BVDV infections. Sixteen colostrum-deprived, BVDV-free, and BVDV-antibody-free calves were infected with 6 different BVDV strains. Clinical signs, seroconversion, and virus isolation indicated that inoculated animals did replicate virus. Virus could be detected in 19% (3/16) of acutely infected animals by the RT-PCR/probe technique. No acutely infected animals were positive by IHC.

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Validation of real-time polymerase chain reaction versus conventional polymerase chain reaction for diagnosis of brucellosis in cattle sera

Veterinary World ◽

10.14202/vetworld.2021.144-154 ◽

2021 ◽

Vol 14 (1) ◽

pp. 144-154

Author(s):

Nour H. Abdel-Hamid ◽

Eman I. M. Beleta ◽

Mohamed A. Kelany ◽

Rania I. Ismail ◽

Nadia A. Shalaby ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Roc Curve ◽

Diagnostic Performance ◽

Rt Pcr ◽

Conventional Pcr ◽

Chain Reaction ◽

Serum Samples ◽

Polymerase Chain ◽

Pcr Targeting ◽

Pcr Techniques

Background and Aim: Different polymerase chain reaction (PCR) techniques have and are still being used for the direct detection of Brucella DNA in serum samples of different animal species and humans without being validated or properly validated, resulting in discrepancies. Thus, this study aimed to evaluate the diagnostic performance of the TaqMan Real- Time-PCR (RT-PCR) targeting the bcsp31 gene versus conventional PCR for the accurate diagnosis of brucellosis at the genus level in cattle sera. Materials and Methods: One hundred and eighty-four serum samples were collected from bacteriologically positive and negative cows with ages ranging from 1 to 5 years old at some infected private farms in the Nile Delta under quarantine measures as well as brucellosis free farms. These samples were classified into four groups after serological diagnosis and investigated by TaqMan RT-PCR and conventional PCR targeting the IS711 gene for Brucella DNA detection. The diagnostic performance characteristics of both PCR techniques were estimated considering the bacteriological results as a gold standard. Results: TaqMan RT-PCR revealed superiority over conventional PCR; it was able to detect Brucella DNA in 95% (67/70) and 89% (25/28) of the cattle sera samples belonging to Group 1 (serologically and bacteriologically positive) and Group 2 (serologically negative but bacteriologically positive), respectively. On evaluating the diagnostic performance, TaqMan RT-PCR showed superior diagnostic sensitivity (93.9%), diagnostic specificity (88.4%), performance index (182.3), almost perfect kappa agreement (0.825±0.042), strong positive correlation (r=0.826), high accuracy based on the receiver operating characteristic (ROC) curve, and area under the ROC curve (0.911) at p<0.05 and CI of 95%. Conclusion: A cattle serum sample is not the metric of choice for targeting Brucella genomic DNA by conventional PCR. The time-saving and rapid TaqMan RT-PCR method revealed a better diagnostic performance in the detection of Brucella DNA in cattle sera. Such performance offered by TaqMan RT-PCR may be considered a step toward the possibility of using such technology in the direct differentiation between Brucella-infected and -vaccinated cattle immunized by smooth vaccines from cattle sera using primers specific for such vaccines.

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

Acta veterinaria ◽

10.1515/acve-2015-0047 ◽

2015 ◽

Vol 65 (4) ◽

pp. 557-567 ◽

Cited By ~ 2

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.

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

10.1101/2020.08.24.264853 ◽

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.

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Seroprevalence of rickettsia spp. and a study of the tick fauna in dogs from the municipality of Seropédica, State of Rio de Janeiro

Semina Ciências Agrárias ◽

10.5433/1679-0359.2015v36n6p3787 ◽

2015 ◽

Vol 36 (6) ◽

pp. 3787 ◽

Cited By ~ 1

Author(s):

Matheus Dias Cordeiro ◽

Vanessa De Almeida Raia ◽

Adriano Pinter ◽

Nathalie Costa da Cunha ◽

Celso Eduardo de Souza ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Rio De Janeiro ◽

Spotted Fever ◽

Rhipicephalus Sanguineus ◽

Spotted Fever Group ◽

Chain Reaction ◽

Serum Samples ◽

Crude Antigen ◽

Polymerase Chain ◽

Time Of Collection

The aim of this study was to investigate the presence of anti-Rickettsia spp. antibodies, the tick fauna, and the ticks that are carriers of rickettsiae of the spotted fever group (SFG). About 68 (24%) of the 283 serum samples tested by indirect immunofluorescence (IFA) reacted against the R. rickettsii crude antigen. The titers varied between 1:64 and 1:512. At the time of collection, 189 (64.5%) of the 293 dogs included in this study, were infested with ticks. Ticks classified as Rhipicephalus sanguineus and Amblyomma sculptum were identified. None of the ticks examined for SFG rickettsiae using polymerase chain reaction (PCR) were positive. The presence of the anti-R. rickettsii antibodies detected by IFA, albeit at low titers, suggests the circulation of SFG rickettsiae, which requires permanent surveillance because there are records on human fatalities related to spotted fever and to avoid any future threats to the students moving extensively in the areas near of the Rural Federal University of Rio de Janeiro.

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DETEKSI VIRUS JAPANESE ENCEPHALITIS PADA MANUSIA DAN VEKTOR DI KABUPATEN TULUNGAGUNG, JAWA TIMUR

Vektora Jurnal Vektor dan Reservoir Penyakit ◽

10.22435/vk.v12i1.2649 ◽

2020 ◽

Vol 12 (1) ◽

pp. 33-40

Author(s):

Nova Pramestuti ◽

Tri Wijayanti ◽

Dyah Widiastuti ◽

Tri Isnani

Keyword(s):

Polymerase Chain Reaction ◽

Human Serum ◽

Japanese Encephalitis ◽

Immunosorbent Assay ◽

Culex Quinquefasciatus ◽

Rt Pcr ◽

Chain Reaction ◽

Serum Samples ◽

Polymerase Chain ◽

Anopheles Vagus

Japanese encephalitis is a zoonotic disease transmitted by mosquitoes with pigs as the main reservoir. A small percentage of infected people experience acute encephalitis syndrome, with one in four cases fatal. Recently, the existence of a growing pig population has the potential to increase the risk of JE transmission in Tulung Agung, East Java, Indonesia. The purpose of this study was to detect JE infection in humans and mosquitoes in Tulungagung Regency. A cross-sectional design was conducted from March to October 2016. Human blood screening was carried out in six hospitals in Tulungagung and the mosquitos survey was carried out using human landing collection, resting collection, and light trap around the pig farms in Kaliwungu District. Detection of JE infection was carried out by indirect immuno-linked immunosorbent assay (ELISA) testing of IgM/IgG in human serum samples and Polymerase Chain Reaction (RT-PCR). Transcription reaction in mosquitoes. The ELISA test showed that one of 19 human serum samples was confirmed positive with JE specific IgG. The result of the mosquito survey showed that Anopheles vagus was predominantly collected in Kaliwungu village, while Culex quinquefasciatus was was predominantly species collected in Pulosari Village. The analyses using molecular assays showed that all captured mosquitoes were negative Javanese encephalitis virus (JEV). Abstrak Japanese Encephalitis (JE) merupakan penyakit zoonotik yang ditularkan oleh nyamuk dengan reservoir utama babi. Sebagian kecil orang yang terinfeksi mengalami radang otak (ensefalitis), sekitar satu dari empat kasus berakibat fatal. Peningkatan populasi ternak babi di Kabupaten Tulungagung berpotensi menyebarkan virus JE. Tujuan penelitian untuk mendeteksi infeksi JE pada manusia dan nyamuk di Kabupaten Tulungagung. Penelitian dilakukan pada Bulan Maret-Oktober tahun 2016 dengan desain studi potong lintang. Survey darah manusia dilakukan pada enam rumah sakit di Kabupaten Tulungagung. Survei nyamuk dilakukan satu kali dengan metode umpan badan manusia dan perangkap cahaya, serta penangkapan nyamuk istirahat di sekitar peternakan babi di Kecamatan Kaliwungu. Deteksi infeksi JE dilakukan dengan pemeriksaan Indirect Enzyme-Linked Immunosorbent Assay (ELISA) IgM/IgG pada sampel serum manusia dan Reverse Transcription Polymerase Chain Reaction (RT-PCR) pada nyamuk. Hasil pemeriksaan laboratorium terhadap antibodi IgM/IgG JE menunjukkan satu kasus positif IgG JE dari 19 sampel serum manusia yang diperiksa. Spesies nyamuk yang tertangkap di Desa Kaliwungu didominasi Anopheles vagus, sedangkan di Desa Pulosari didominasi Culex quinquefasciatus. Hasil pemeriksaan RT-PCR terhadap semua sampel nyamuk yang tertangkap menunjukkan negatif virus JE. Satu pasien ditemukan positif antibodi IgG Japanese encephalitis di Kabupaten Tulungagung.

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RBD-IgG levels correlate with protection in Residents Facing SARS-CoV-2 B.1.1.7 Outbreaks

10.22541/au.163254719.96929295/v1 ◽

2021 ◽

Author(s):

Hubert Blain ◽

Edouard Tuaillon ◽

Lucie Gamon ◽

Amandine Pisoni ◽

Stephanie Miot ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Humoral Response ◽

Nasopharyngeal Swab ◽

Receptor Binding Domain ◽

Limited Information ◽

Rt Pcr ◽

Chain Reaction ◽

Neutralization Activity ◽

Polymerase Chain

Background Limited information exists on nursing home (NH) residents regarding BNT162b2/Pfizer vaccine efficacy in preventing SARS-CoV-2 and severe Covid-19, and its association with post-vaccine humoral response. Methods 396 residents from seven NHs suffering a SARS-CoV-2 B.1.1.7 (VOC-α) outbreak at least 14 days after a vaccine campaign were repeatedly tested using SARS-CoV-2 real-time reverse-transcriptase polymerase chain reaction on nasopharyngeal swab test (RT-PCR). SARS-CoV-2 Receptor-Binding Domain (RBD) of the S1 subunit (RBD-IgG) was measured in all residents. Nucleocapsid antigenemia (N-Ag) was measured in RT-PCR-positive residents, and serum neutralizing antibodies in vaccinated residents from one NH. Results The incidence of positive RT-PCR was lower in residents vaccinated by two doses (22.7%) vs one dose (32.3%) or non-vaccinated residents (43.7%)(p<0.01). Covid-19-induced deaths were observed in 10.4% of the non-vaccinated residents, in 6.4% of those who had received one dose, and in 0.9% with two doses (p=0.0007). Severe symptoms were more common in infected non-vaccinated (21.0%) vs vaccinated residents (47.6%, p=0.002). Higher levels of RBD-IgG (n=325) were associated with a lower SARS-CoV-2 incidence. No in vitro serum neutralization activity was found for RBD-IgG levels below 1,050 AU/mL. RBD-IgG levels were inversely associated with N-Ag levels, found as a risk factor of severe Covid-19. Conclusions Two BNT162b2/Pfizer doses are associated with a 48% reduction of SARS-CoV-2 incidence and a 91.3% reduction of death risk in residents from NHs facing a VOC-α outbreak. BNT162b2/Pfizer efficacy was partly predicted by post-vaccine RBD-IgG levels.

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

The Journal of Urology ◽

10.1016/s0022-5347(18)33708-x ◽

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

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

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648955 ◽

1994 ◽

Vol 72 (05) ◽

pp. 762-769 ◽

Cited By ~ 11

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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