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

Jing, Wang; Luo, Yakun; Liang, Lin; Li, Jinxiang; Cui, Shangjin

doi:10.1007/s00705-018-3982-8

Cited by 14 publications

(12 citation statements)

References 22 publications

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“…All samples were stored at −80 • C to prevent virus deactivation. The infection status and co-infections were assessed by investigating the presence of canine parvovirus 2 (CPV-2), canine distemper virus (CDV), CAdV-1, CAdV-2, canine coronavirus (CRCoV), Canine Circovirus (CACV), and canine bocavirus (CBOV) using polymerase chain reaction (PCR) or real-time PCR, as documented in previous reports (23)(24)(25)(26). The positive infection rate (%) was calculated as follows: Positive infection rate of samples (%) = (number of positive samples/total number of samples) × 100.…”

Section: Clinical Sample Collection and Virus Screeningmentioning

confidence: 99%

“…Based on the composition of the fibrillar structural functional regions of human adenoviruses, we found that the structural functional regions of CAdV-2 are composed of amino acids 1-42, 43-365, and 366-542, which are located in the tail, shaft, and head regions, respectively (28)(29)(30). The mutant amino acids in the 19 CAdV-2 strains were located at sites 18,25,26,30,36,37,40, and 41, which were located in the tail of the fibrillar process; sites 44, 45, 50, etc., which were located on the basal axis of the fibrillar process, also known as the shaft; and sites 370, 374, 393, etc., which were located in the ganglion region of Fiber, also known as the knob (Figure 4A). There are four differences between Figures 4B,C at sites 37, 77, 408, 520, 521, 523-532 of Fiber.…”

Section: Protein Structure Predictionmentioning

confidence: 99%

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Novel Genotype Definition and the First Epidemiological Investigation of Canine Adenovirus Type 2 in Dogs in Central China

Ji¹,

Li²,

Hu³

et al. 2020

Front. Vet. Sci.

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Infections caused by canine adenovirus (CAdV) type 1 have been reported worldwide in the past two decades. However, only few studies have specifically reported the prevalence of CAdV type 2 (CAdV-2). The present study investigated the persistent circulation of CAdV-2 in dogs with diarrhea in the Henan, Hubei, and Jiangsu provinces in central China from 2017 to 2019. We conducted polymerase chain reaction for detecting CAdV-2 and other related pathogens in 224 rectal swabs of pet dogs and the co-infection of canine diseases was also analyzed. In addition, the structural protein genes-Fiber, Hexon, and Penton-of the isolated CAdV-2 strains were sequenced and analyzed. The similarity between Hexon and Penton among the 19 strains was 97.4%, as revealed by sequence alignment. Multiple sequence alignment results showed that the Fiber gene sequences of these CAdV-2 strains shared 97.4-99.8% nucleotide and 94.1-99.3% amino acid identity with reference sequences and shared only 79.0-80.5% nucleotide and 77.3-80.5% amino acid identity with the vaccine strain CLL, indicating that Fiber harbored most of the variant sites. Furthermore, pairwise sequence comparisons of Hexon of CH-JS-1901 and CH-HN-1801 with that of India2006 revealed a novel genotype. Furthermore, protein model prediction showed that the amino acid mutation of fiber protein in 19 strains was located in the head region, that may cause structural changes on the surface of the fiber protein. These findings are of significance for monitoring the epidemiology of CAdV-2 infection and developing a novel vaccine which contribute to understanding genetic evolution of CAdV-2 in China.

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Section: Clinical Sample Collection and Virus Screeningmentioning

confidence: 99%

Section: Protein Structure Predictionmentioning

confidence: 99%

Novel Genotype Definition and the First Epidemiological Investigation of Canine Adenovirus Type 2 in Dogs in Central China

Ji¹,

Li²,

Hu³

et al. 2020

Front. Vet. Sci.

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“…At present, detection of CDV infection has mainly relied on RT-PCR or real-time RT-PCR and testing the CDV viral particles using immune colloidal gold test strips. However, RT-PCR assays can only be carried out in well-equipped laboratories (Kim et al 2006;Wang et al 2018), while immune colloidal gold test strip testing is expensive and therefore unsuitable for testing of large numbers of samples. For these reasons, here we developed a sandwich enzyme-linked immunosorbent assay (ELISA) as a sensitive, specific, and easy-to-implement assay for detection of viral antigens that could be suitable for high-throughput testing applications.…”

Section: Introductionmentioning

confidence: 99%

Development of a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay for detecting canine distemper virus

Yuan

Zhang

et al. 2020

Appl Microbiol Biotechnol

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Canine distemper virus (CDV) infection causes mass mortality in diverse carnivore species. For effective virus surveillance, rapid and sensitive assays are needed to detect CDV in field samples. In this study, after BABL/c mice were immunized with recombinant CDV-fusion (F) protein, monoclonal antibodies (mAbs) against recombinant CDV-F protein (designated 1A5, 1A6, and 7D5) were produced using traditional hybridoma cell technology. Next, capture antibody (1A6, 800 ng/well) and horseradish peroxidase (HRP)–conjugated detection antibody (HRP-7D5, 1:100, 500 ng/well) were used in a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay (ELISA) for CDV detection after optimization of both mAb amounts per well using a checkerboard titration test. Based on sandwich ELISA test results for 120 known CDV-negative samples, the cutoff value for a positive result was set to an OD450 nm value ≥ 0.196. As compared with test results obtained from commercial immune colloidal gold test strips, the low limits of detection for the two assays were revealed to be 100 TCID50 per 100 μL. In addition, the sandwich ELISA agreed 100% and 96.4% with commercial immune colloidal gold test strips when testing serum and stool samples. The sandwich ELISA assay provided statistically similar CDV detection. Thus, the sandwich ELISA developed here to detect CDV in fecal and serum samples provided good sensitivity, high specificity, and good reproducibility and should serve as an ideal method for large-scale surveillance of CDV infections in carnivores. Key points • Three CDV mAbs that recognized different epitopes and bound to virion were generated. • The sandwich ELISA based mAbs to detect CDV in fecal and serum samples was developed. • The sandwich ELISA is an ideal method for detecting CDV infections in the field.

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“…Canine distemper is a contagious disease with multisystem infection caused by canine distemper virus (CDV), which affects the health of multiple species (1,2). The disease is characterized by high fever, diarrhea, encephalitis, immunosuppression, and other symptoms.…”

Section: Introductionmentioning

confidence: 99%

Identification of Linear B-Cell Epitopes on Hemagglutinin Protein of Canine Distemper Virus Using Two Monoclonal Antibodies

et al. 2020

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Canine distemper virus (CDV) belongs to the Morbillivirus genus of the Paramyxoviridae family, which causes a threat to the domestic dog and fur-animal industry. Hemagglutinin protein is a major membrane protein of the vital molecular factor in CDV tropism, also known to induce hosts to produce neutralizing antibodies. In the current study, we prepared two monoclonal antibodies, 1A5 and 2B8, against the H protein of the CDV-PS strain. A series of partially overlapping synthetic peptides covering the hemagglutinin protein (amino acids 50-204) were screened to define the linear epitope identified by 1A5 and 2B8 mAbs. 120 QKTNFFNPNREFDFR 134 (F8) and 178 ARGDIFPPY 186 (F14-1) are minimal linear epitopes recognized by 1A5 and 2B8 mAbs, respectively. Further investigations revealed that F8 is conserved in different CDV strains; however, F14-1 contains mutant residues 178, 179, and 180. The epitopes F8 and F14-1 localized at the surface of hemagglutinin protein in a three-dimensional (3D) structure. CDV-infected dog serum can also recognize the identified B-cell epitopes.

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

Cited by 14 publications

References 22 publications

Novel Genotype Definition and the First Epidemiological Investigation of Canine Adenovirus Type 2 in Dogs in Central China

Novel Genotype Definition and the First Epidemiological Investigation of Canine Adenovirus Type 2 in Dogs in Central China

Development of a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay for detecting canine distemper virus

Identification of Linear B-Cell Epitopes on Hemagglutinin Protein of Canine Distemper Virus Using Two Monoclonal Antibodies

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