scholarly journals A New Molecular Detection System for Canine Distemper Virus Based on a Double-Check Strategy

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1632
Author(s):  
Sabrina Halecker ◽  
Sabine Bock ◽  
Martin Beer ◽  
Bernd Hoffmann
Keyword(s):  
Canine Distemper Virus ◽  
Parallel Implementation ◽  
Detection System ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Wild Animals ◽  
Canine Distemper ◽  
One Step ◽  
Diagnostic Sensitivity And Specificity

Due to changing distemper issues worldwide and to inadequate results of an inter-laboratory study in Germany, it seems sensible to adapt and optimize the diagnostic methods for the detection of the canine distemper virus (CDV) to the new genetic diversity of virus strains. The goal of the project was the development, establishment and validation of two independent one-step reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) methods for the safe detection of CDV in domestic and wild animals. For this purpose, an existing CDV-RT-qPCR was decisively adapted and, in addition, a completely new system was developed. Both CDV-RT-qPCR systems are characterized by a very high, comparable analytical and diagnostic sensitivity and specificity and can be mutually combined with inhibition or extraction controls. The reduction in the master mix used allows for the parallel implementation of both CDV-RT-qPCR systems without significant cost increases. For validation of the new CDV-RT-qPCR duplex assays, a panel comprising 378 samples derived from Germany, several European countries and one African country were tested. A sensitivity of 98.9% and a specificity of 100% were computed for the new assays, thus being a reliable molecular diagnostic tool for the detection of CDV in domestic and wild animals.

scholarly journals Canine distemper virus detection by different methods of One-Step RT-qPCR

2016 ◽  
Vol 46 (9) ◽  
pp. 1601-1606
Author(s):  
Claudia de Camargo Tozato ◽  
Vívian Ferreira Zadra ◽  
Caroline Rodrigues Basso ◽  
João Pessoa Araújo Junior
Keyword(s):  
Urine Sample ◽  
Canine Distemper Virus ◽  
Limit Of Detection ◽  
Analytical Sensitivity ◽  
Canine Distemper ◽  
System A ◽  
Virus Rna ◽  
One Step ◽  
Commercial Kits ◽  
The One

ABSTRACT: Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Real-time RT-PCR (RT-qPCR) assays were tested and compared for analytical sensitivity to Canine Distemper Virus RNA detection: a One-Step RT-qPCR (system A) and a One-Step RT-qPCR combined with NESTED-qPCR (system B). Limits of detection for both systems were determined using a serial dilution of Canine Distemper Virus synthetic RNA or a positive urine sample. In addition, the same urine sample was tested using samples with prior centrifugation or ultracentrifugation. Commercial kits of One-Step RT-qPCR assays detected canine distemper virus RNA in 10 (100%) urine samples from symptomatic animals tested. The One-Step RT-qPCR kit that showed better results was used to evaluate the analytical sensitivity of the A and B systems. Limit of detection using synthetic RNA for the system A was 11 RNA copies µL-1 and 110 RNA copies µl-1 for first round System B. The second round of the NESTED-qPCR for System B had a limit of detection of 11 copies µl-1. Relationship between Ct values and RNA concentration was linear. The RNA extracted from the urine dilutions was detected in dilutions of 10-3 and10-2 by System A and B respectively. Urine centrifugation increased the analytical sensitivity of the test and proved to be useful for routine diagnostics. The One-Step RT-qPCR is a fast, sensitive and specific method for canine distemper routine diagnosis and research projects that require sensitive and quantitative methodology.

scholarly journals A fast and simple one-step duplex PCR assay for canine distemper virus (CDV) and canine coronavirus (CCoV) detection

2018 ◽  
Vol 163 (12) ◽  
pp. 3345-3349 ◽  
Author(s):  
Jing Wang ◽  
Yakun Luo ◽  
Lin Liang ◽  
Jinxiang Li ◽  
Shangjin Cui
Keyword(s):  
Canine Distemper Virus ◽  
Duplex Pcr ◽  
Canine Distemper ◽  
Pcr Assay ◽  
Canine Coronavirus ◽  
One Step ◽  
Duplex Pcr Assay

scholarly journals An Outbreak of Canine Distemper Virus in Tigers (Panthera tigris): Possible Transmission from Wild Animals to Zoo Animals

2012 ◽  
Vol 74 (6) ◽  
pp. 699-705 ◽  
Author(s):  
Yumiko NAGAO ◽  
Yohei NISHIO ◽  
Hiroshi SHIOMODA ◽  
Seiji TAMARU ◽  
Masayuki SHIMOJIMA ◽  
...  
Keyword(s):  
Canine Distemper Virus ◽  
Panthera Tigris ◽  
Wild Animals ◽  
Canine Distemper ◽  
Zoo Animals

scholarly journals Immunopathogenic and Neurological Mechanisms of Canine Distemper Virus

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Otávio Valério Carvalho ◽  
Clarisse Vieira Botelho ◽  
Caroline Gracielle Torres Ferreira ◽  
Paulo Oldemar Scherer ◽  
Jamária Adriana Pinheiro Soares-Martins ◽  
...  
Keyword(s):  
Canine Distemper Virus ◽  
Clinical Manifestations ◽  
Specific Treatment ◽  
Viral Disease ◽  
Diagnostic Methods ◽  
Lymphoid Tissues ◽  
Canine Distemper ◽  
Worldwide Distribution ◽  
Natural Hosts ◽  
The Central Nervous System

Canine distemper is a highly contagious viral disease caused by the canine distemper virus (CDV), which is a member of theMorbillivirusgenus, Paramyxoviridae family. Animals that most commonly suffer from this disease belong to the Canidae family; however, the spectrum of natural hosts for CDV also includes several other families of the order Carnivora. The infectious disease presents worldwide distribution and maintains a high incidence and high levels of lethality, despite the availability of effective vaccines, and no specific treatment. CDV infection in dogs is characterized by the presentation of systemic and/or neurological courses, and viral persistence in some organs, including the central nervous system (CNS) and lymphoid tissues. An elucidation of the pathogenic mechanisms involved in canine distemper disease will lead to a better understanding of the injuries and clinical manifestations caused by CDV. Ultimately, further insight about this disease will enable the improvement of diagnostic methods as well as therapeutic studies.

scholarly journals OmniSARS2: A Highly Sensitive and Specific RT-qPCR-Based COVID-19 Diagnostic Method Designed to Withstand SARS-CoV-2 Lineage Evolution

Biomedicines ◽  
2021 ◽  
Vol 9 (10) ◽  
pp. 1314
Author(s):  
Eduarda Carvalho-Correia ◽  
Carla Calçada ◽  
Fernando Branca ◽  
Nuria Estévez-Gómez ◽  
Loretta De Chiara ◽  
...  
Keyword(s):  
Cross Reactivity ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
Analytical Sensitivity ◽  
S Gene ◽  
Genomic Knowledge ◽  
Gene Assay ◽  
One Step

Extensive transmission of SARS-CoV-2 during the COVID-19 pandemic allowed the generation of thousands of mutations within its genome. While several of these become rare, others largely increase in prevalence, potentially jeopardizing the sensitivity of PCR-based diagnostics. Taking advantage of SARS-CoV-2 genomic knowledge, we designed a one-step probe-based multiplex RT-qPCR (OmniSARS2) to simultaneously detect short fragments of the SARS-CoV-2 genome in ORF1ab, E gene and S gene. Comparative genomics of the most common SARS-CoV-2 lineages, other human betacoronavirus and alphacoronavirus, was the basis for this design, targeting both highly conserved regions across SARS-CoV-2 lineages and variable or absent in other Coronaviridae viruses. The highest analytical sensitivity of this method for SARS-CoV-2 detection was 94.2 copies/mL at 95% detection probability (~1 copy per total reaction volume) for the S gene assay, matching the most sensitive available methods. In vitro specificity tests, performed using reference strains, showed no cross-reactivity with other human coronavirus or common pathogens. The method was compared with commercially available methods and detected the virus in clinical samples encompassing different SARS-CoV-2 lineages, including B.1, B.1.1, B.1.177 or B.1.1.7 and rarer lineages. OmniSARS2 revealed a sensitive and specific viral detection method that is less likely to be affected by lineage evolution oligonucleotide–sample mismatch, of relevance to ensure the accuracy of COVID-19 molecular diagnostic methods.

scholarly journals One-step triplex PCR/RT-PCR to detect canine distemper virus, canine parvovirus and canine kobuvirus

2019 ◽  
Vol 81 (7) ◽  
pp. 1040-1042 ◽  
Author(s):  
Dafei LIU ◽  
Fei LIU ◽  
Dongchun GUO ◽  
Xiaoliang HU ◽  
Zhijie LI ◽  
...  
Keyword(s):  
Canine Distemper Virus ◽  
Canine Parvovirus ◽  
Canine Distemper ◽  
Rt Pcr ◽  
One Step ◽  
Canine Kobuvirus

scholarly journals Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR

2001 ◽  
Vol 2 (1) ◽  
pp. 59 ◽  
Author(s):  
Yong Hwan Kim ◽  
Kyu Woan Cho ◽  
Hwa Young Youn ◽  
Han Sang Yoo ◽  
Hong Ryul Han
Keyword(s):  
Nested Pcr ◽  
Canine Distemper Virus ◽  
Canine Distemper ◽  
Rt Pcr ◽  
One Step

scholarly journals Death comes for us all: an interplay of habitat selection, movement, and social behavior relate to cause specific mortality among grey wolves

2021 ◽  
Author(s):  
Julie W. Turner ◽  
Christina M. Prokopenko ◽  
Katrien A. Kingdon ◽  
Daniel L. J. Dupont ◽  
Sana Zabihi-Seissan ◽  
...  
Keyword(s):  
Social Behavior ◽  
Habitat Selection ◽  
Canine Distemper Virus ◽  
Biological Processes ◽  
Wild Animals ◽  
Disease Dynamics ◽  
Canine Distemper ◽  
Mortality Risks ◽  
Specific Mortality ◽  
Step Selection

AbstractAvoiding death infects biological processes, including behavior. Habitat selection, movement, and sociality are highly flexible behaviors that influence the mortality risks and subsequent fitness of individuals. In the Anthropocene, animals are experiencing increased risks from direct human causes and increased spread of infectious diseases. Using integrated step selection analysis, we tested how the habitat selection, movement, and social behaviors of grey wolves vary as an individual dies due to humans or canine distemper virus (CDV) and how those behaviors may vary in the lead up to death. Behaviors that changed prior to death were strongly related to how an animal eventually died. Wolves killed by humans moved slower than wolves that survived and selected to be nearer roads closer in time to their death. Wolves that died due to CDV moved progressively slower as they neared death and reduced their avoidance of wet habitats. All animals, regardless of dying or not maintained strong selection to be near packmates across time, which seemingly contributed to disease dynamics in the packs that became infected with CDV. Habitat selection, movement, and sociality interact to put individuals and groups at greater risks, influencing their cause-specific mortality.Lay SummaryNot much is known about behaviors prior to death in wild animals. Grey wolves killed by humans selected to be in riskier areas increasingly prior to their deaths. Wolves that died due to disease moved slower and changed their habitat selection to be in areas with more water as they became sicker. Sick wolves also continued to select for packmates, increasing the chances that the whole pack would succumb to the disease.

Detection of Canine Distemper Virus Nucleoprotein RNA by Reverse Transcription-PCR Using Serum, Whole Blood, and Cerebrospinal Fluid from Dogs with Distemper

1999 ◽  
Vol 37 (11) ◽  
pp. 3634-3643 ◽  
Author(s):  
A. L. Frisk ◽  
M. König ◽  
A. Moritz ◽  
W. Baumgärtner
Keyword(s):  
Cerebrospinal Fluid ◽  
Reverse Transcription ◽  
Whole Blood ◽  
Canine Distemper Virus ◽  
Detection System ◽  
Nucleotide Sequence Analysis ◽  
Restriction Enzyme Digestion ◽  
Canine Distemper ◽  
Rt Pcr ◽  
Reverse Transcription Pcr

Reverse transcription-PCR (RT-PCR) was used to detect canine distemper virus (CDV) nucleoprotein (NP) RNA in serum, whole blood, and cerebrospinal fluid (CSF) samples from 38 dogs with clinically suspected distemper. Results were correlated to clinical findings, anti-CDV neutralizing antibody titers, postmortem findings, and demonstration of CDV NP antigen by immunohistochemistry. The specificity of the RT-PCR was ensured by amplification of RNA from various laboratory CDV strains, restriction enzyme digestion, and Southern blot hybridization. In 29 of 38 dogs, CDV infection was confirmed by postmortem examination and immunohistochemistry. The animals displayed the catarrhal, systemic, and nervous forms of distemper. Seventeen samples (serum, whole blood, or CSF) from dogs with distemper were tested with three sets of primers targeted to different regions of the NP gene of the CDV Onderstepoort strain. Expected amplicons were observed in 82, 53, and 41% of the 17 samples, depending upon the primer pair used. With the most sensitive primer pair (primer pair I), CDV NP RNA was detected in 25 of 29 (86%) serum samples and 14 of 16 (88%) whole blood and CSF samples from dogs with distemper but not in body fluids from immunohistochemically negative dogs. Nucleotide sequence analysis of five RT-PCR amplicons from isolates from the field revealed few silent point mutations. These isolates exhibited greater homology to the Rockborn (97 to 99%) than to the Onderstepoort (95 to 96%) CDV strain. In summary, although the sensitivity of the RT-PCR for detection of CDV is strongly influenced by the location of the selected primers, this nucleic acid detection system represents a highly specific and sensitive method for the antemortem diagnosis of distemper in dogs, regardless of the form of distemper, humoral immune response, and viral antigen distribution.

Serological and Nucleocapsid Gene Based Molecular Characterization of Canine Distemper Virus (CDV) Isolated from Dogs of Southern Gujarat, India

2020 ◽  
Author(s):  
Dhruv Desai ◽  
Irshadullakhan Kalyani ◽  
Jayesh Solanki ◽  
Dharmesh Patel ◽  
Pushpa Makwana ◽  
...  
Keyword(s):  
Cell Line ◽  
Mdck Cell ◽  
Canine Distemper Virus ◽  
Age Group ◽  
Canine Distemper ◽  
Rt Pcr ◽  
Serum Samples ◽  
Mdck Cell Line ◽  
One Step

Background: The present study was undertaken to diagnose and characterize canine distemper virus (CDV) isolated from dogs of southern Gujarat, India. CDV is lethal disease of canines and felines. Total of 40 different samples were collected from 18 suspected stray dogs having different clinical signs which were processed for diagnosis and characterization of CDV.Methods: All samples were processed by employing different methods like, Immunochromatography based lateral flow test (LFA), IgG based indirect enzyme linked immunosorbent assay (i-ELISA), one step RT-PCR, nested one step RT-PCR and virus isolation in MDCK cell line. Restriction endonuclease (RE) analysis was used to characterize CDV Nucleocapsid (N) gene. Conclusion: Only 04 samples (02 nasal and 02 ocular swabs) of 02 dogs found positive for LFA, while 14 serum samples out of 17 samples of 18 dogs found positive for IgG antibody. As all dogs were unvaccinated, serum samples found positive in IgG based ELISA considered for confirmative positive for CDV infection. Whereas 13 samples of 10 dogs found positive for one step RT-PCR and nested one step RT-PCR. In RE digestion, characteristic two bands were found. All representative CDV positive samples of 10 dogs showed characteristic cytopathic effect in MDCK cell line. On age group wise percent positivity was found 71.42 % (05/07) in 0 - ≤6 months, while 77.77% (07/09) in 6- ≤12 months of age group, whereas, both samples were found positive in 12 months and above group. Overall 77.77% (14/18) dogs found positive for CDV infection. To the best of our knowledge, this is the first report on study of CDV infection in dogs from Gujarat state, India.

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