Detection of canine adenoviral infections in urine and faeces by the polymerase chain reaction

Chaturvedi, Uttara; Tiwari, Ashok Kumar; Ratta, Barkha; Ravindra, P. V.; Rajawat, Yogendra Singh; Palia, Sudesh Kumar; A, Rai

doi:10.1016/j.jviromet.2008.01.024

Cited by 29 publications

(28 citation statements)

References 11 publications

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“…Viral DNA was extracted from serum and rectal swabs using the DNA Extract Mini Kit (QIAGEN, Hilden, Germany) as per the manufacturer's instructions. A portion of the E3 gene and its flanking regions was amplified using two conserved primers that produce different length fragments to differentiate between the two adenovirus types: 508 bp for CAdV-1 and 1030 bp for CAdV-2 (Hu et al, 2001;Chaturvedi et al, 2008). Thirty cycles were performed as follows: 95 • C for 5 min, 94 • C for 40 s, 58 • C for 1 min, 72 • C for 1 min, and 72 • C for 10 min.…”

Section: Clinical Application Of the Icsmentioning

confidence: 99%

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Wang

Wen

et al. 2019

Front. Microbiol.

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Although canine adenovirus (CAdV) is highly prevalent in dogs, there is currently a lack of a quick diagnostic method. In this study, we developed a rapid immunochromatographic strip (ICS) assay using colloidal gold coupled to CAdV-2-specific monoclonal antibodies (mAbs). BALB/c mice were immunized with a purified CAdV-2 suspension, and four mAbs (belonging to two different epitopes) were generated and designated as 2C1, 7D7, 10D1, and 4G1. Western blot and protein spectral analysis indicated that the hexon protein of CAdV-2 recognized all four mAbs. The colloidal gold-coupled 7D7 and 2C1 mAbs were chosen for inclusion in the rapid ICS assay. The optimal concentrations of the coating antibody (2C1), the capture antibody (7D7), and the goat anti-mouse antibody were 1.0 mg/ml, 10 µg/ml, and 2.0 mg/ml, respectively. The limit of detection was approximately 2.0 × 10 2 tissue culture infective dose (TCID 50 )/ml. Other common canine viruses were tested to evaluate the specificity of the ICS, and positive results were observed for only CAdV-1 and CAdV-2. The ICS test was conducted on 360 samples to detect CAdV, and the results were compared with those of polymerase chain reaction (PCR) tests. The ICS test was found to be a sufficiently sensitive and specific detection method for the convenient and rapid detection of CAdV.

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Section: Clinical Application Of the Icsmentioning

confidence: 99%

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Wang

Wen

et al. 2019

Front. Microbiol.

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“…The results obtained from PCR (Chaturvedi et al, 2008;Hu et al, 2001), direct nucleotide sequencing, and melting curve analysis of the two CAdV types were in complete agreement. Thus, the results confirm that the MCA-based SYBR Green qPCR performed under the optimized conditions is reproducible and highly efficient and specific in discriminating between CAdV-1 and CAdV-2.…”

Section: Discussionmentioning

confidence: 60%

“…Field viruses were previously identified in our laboratory by testing different biological matrices and using a PCR assay we were able to differentiate between the two adenovirus types (Chaturvedi et al, 2008;Hu et al, 2001). All the obtained PCR products were sequenced to confirm the CAdV typing.…”

Section: Methodsmentioning

confidence: 92%

“…are mostly unable to distinguish the two viruses, and also for the widespread use of a modified live CAdV-2 vaccine. The molecular biology-based methods are more reliable but, to date, only one PCR assay able to distinguish the two CAdV types was developed (Hu et al, 2001;Chaturvedi et al, 2008). In order to reduce the time of execution and, at the same time, to increase the sensitivity of the molecular diagnosis, a SYBR Green real-time PCR (qPCR) assay was developed that is able to detect and to distinguish between CAdV-1 and CAdV-2 via a melting curve analysis (MCA).…”

Section: Introductionmentioning

confidence: 99%

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Development of a SYBR Green real-time PCR assay with melting curve analysis for simultaneous detection and differentiation of canine adenovirus type 1 and type 2

Balboni¹,

Dondi²,

Prosperi³

et al. 2015

Journal of Virological Methods

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Canine adenovirus type 1 (CAdV-1) and canine adenovirus type 2 (CAdV-2) cause infectious canine hepatitis (ICH) and infectious tracheobronchitis (ITB) in dogs, respectively. Cases of ICH have been documented in recent years and recent surveys have demonstrated a wide percentage of asymptomatic CAdV-1 infection in the canine population. Since both CAdV types are detectable in the same biological matrices, and viral coinfection with CAdV-1 and CAdV-2 are reported with high frequency, it is urgent to have available a rapid, highly sensitive and specific assay for the diagnosis of CAdV infection and distinction between CAdV-1 and CAdV-2. In order to detect canine adenovirus in biological samples and to rapidly distinguish the two viral types, a SYBR Green real-time PCR assay was optimized to discriminate CAdV-1 and CAdV-2 via a melting curve analysis. The developed assay showed high sensitivity and reproducibility and was highly efficient and specific in discriminating the two CAdV types. This reliable and rapid technique may represent a simple, useful and economic option for simultaneous CAdV types detection, which would be feasible and attractive for all diagnostic laboratories, both for clinical purposes and for epidemiological investigations.

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“…Extraction was carried out according to the manufacturer's instructions. Primers and methods suitable for both CAV-1 and CAV-2 were used for the PCR, as recommended by HU et al, (2001) andCHATURVEDI et al (2008). Consequently, the formation of PCR products in the expected sizes (508 bp and 1030 bp for CAV-1 and CAV-2, respectively) was analysed using DNA gel electrophoresis.…”

Section: Extraction and Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Detection and molecular characterization of canine adenovirus type 2 (CAV-2) in dogs with respiratory tract symptoms in shelters in Turkey

2018

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Canine adenoviruses are agents responsible for two different infections in Canidae. While canine adenovirus type 1 (CAV-1) causes contagious hepatitis (HCC) in dogs, canine adenovirus type 2 (CAV-2) is responsible for infectious laryngotracheitis (ILT). CAV-2, especially in the respiratory tract, leads to an infection that can result in death in young and cohabitant animals. In public housing such as shelters, in addition to opportunistic infections, a disorder defined as canine infectious respiratory disease (CIRD) may also occur frequently. In this study, 155 nasal swabs were collected from dogs in two shelters where cases of respiratory system infections were closely monitored. These samples were tested for CAV-2 using polymerase chain reaction (PCR) with primers designed for the CAV E3 (Early) gene. Positive amplicons were subjected to DNA sequencing. CAV-2 nucleic acids were present in 2.5% (4/155) of the test samples. The phylogenetic assessment of the amplicon sequences revealed a 97.7%-98.9% similarity in the local viruses. The partial sequence analyses of the E3 gene of CAV-2 showed that Turkish and Chinese strains have differences in 9 amino acids. These differences redounded on phylogenetic analyses, and the virus which was considered as a single group, is now subdivided into two subgroups. One subgroup comprises American-European isolates and the other one consists of Turkish and Chinese isolates, so this subdivision can be classified into at least two subgroups, designated China-Turkey and America-Europe. To our knowledge, this is the first study that has examined the possible role of CAV-2 in respiratory system infections in dogs in Turkey, to provide novel and updated information regarding CAV-2.

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Detection of canine adenoviral infections in urine and faeces by the polymerase chain reaction

Cited by 29 publications

References 11 publications

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Development of an Immunochromatographic Strip for Rapid Detection of Canine Adenovirus

Development of a SYBR Green real-time PCR assay with melting curve analysis for simultaneous detection and differentiation of canine adenovirus type 1 and type 2

Detection and molecular characterization of canine adenovirus type 2 (CAV-2) in dogs with respiratory tract symptoms in shelters in Turkey

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