Research Article
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Rapid molecular detection and isolation of canine distemper virus in naturally infected dogs

Year 2023, , 49 - 56, 30.12.2022

Hasbi Sait Saltık , Mehmet Kale

https://doi.org/10.33988/auvfd.846475

Abstract

The canine distemper virus (CDV), which infects dogs and a broad range of animal species, remains a serious concern in Türkiye and across the world. The current study shows that CDV can be detected and isolated rapidly and specifically in naturally infected dogs. Whole blood, nasal swab, ocular swab, rectal swab, and urine samples from 50 stray dogs were used in the study (n = 250). The presence of the CDV genome was confirmed in 105 (42%) samples using one-Step real-time RT-PCR. In total, 39 dogs were diagnosed with CDV infection based on the detection of cytopathic effects in MDCK, which was verified by the fluorescent antibody technique. A total of 12 one-Step real-time RT-PCR negative samples, consisting of 4 rectal swabs and 8 urine samples, were found to be positive by virus isolation. Blood, nasal swab, ocular swab (P<0.01, r = 1), rectal swab (P<0.01, r = 0.844), and urine samples (P<0.01, r = 0.697) all showed positive correlations in the tests for viral genome detection and virus isolation. CPE levels of high 37 (31.62%), medium 26 (22.23%) and low 54 (46.15%) were detected in a total of 117 (46.8%) samples with viral growth in cell culture. The highest CPE levels detected by FAT were for rectal swab and urine samples. In conclusion, the one-step real-time RT-PCR method on rectal swab samples proved to be a very sensitive method for the rapid and reliable CDV detection. Besides, non-modified MDCK can be used to isolate CDV from naturally infected dogs.

Keywords

Canine cell culture distemper fluorescent antibody TaqMan

Supporting Institution

Burdur Mehmet Akif Ersoy Üniversitesi Bilimsel Araştırma ve Projeler Koordinatörlüğü (BAP)

Project Number

0403-DR-16

Thanks

This article was summarized from the PhD thesis by Hasbi Sait SALTIK (Burdur Mehmet Akif Ersoy University, Institute of Health Sciences).

References

  • Anderson DE, Wang LF (2011): New and emerging paramyxoviruses. 435-459. In: SK Samal (Ed), The Biology of Paramyxoviruses. Caister Academic Press, Maryland, USA
  • Appel MJ, Gillespie JH (1972): Canine distemper virus. 1-96. In: SGC Hallauer and K Meyer (Ed), Virology Monographs. Springer, Vienna.
  • Appel MJG, Summers B (1999): Canine distemper: current status. 68-72. In: L Carmichael (Ed), Recent Advances in Canine Infectious Diseases. International Veterinary Information Service (IVIS).
  • Appel MJG, Summers BA (1995): Pathogenicity of morbilliviruses for terrestrial carnivores. Vet Microbiol, 44, 187-191.
  • Arya M, Shergill IS, Williamson M, et al (2005): Basic principles of real-time quantitative PCR. Expert Rev Mol Diagn, 5, 209-219.
  • Beineke A, Puff C, Seehusen F, et al (2009): Pathogenesis and immunopathology of systemic and nervous canine distemper. Vet Immunol Immunopathol, 127, 1-18.
  • Blancou J (2004): Dog distemper: imported into Europe from South America. Hist Med Vet, 29, 35-41.
  • Blixenkrone-Moeller M, Svansson V, Have P, et al (1993): Studies on manifestations of canine distemper virus infection in an urban dog population. Vet Microbiol, 37, 163-173.
  • Calderon MG, Remorini P, Periolo O, et al (2007): Detection by RT-PCR and genetic characterization of canine distemper virus from vaccinated and non-vaccinated dogs in Argentina. Vet Microbiol, 125, 341-349.
  • Chappuis G (1995): Control of canine distemper. Vet Microbiol, 44, 351-358.
  • Demeter Z, Palade EA, Rusvai M (2009): Canine distemper: Still a major concern in Central Europe. Lucr Stiint Univ Stiint Agric Banat Timis Med Vet, 42, 136-150.
  • Di Sabatino D, Savini G, Lorusso A (2015): Canine distemper and endangered wildlife: Is it time for mandatory vaccination of dogs. Vaccine, 33, 6519.
  • Diaz NM, Mendez GS, Grijalva CJ, et al (2016): Dog overpopulation and burden of exposure to canine distemper virus and other pathogens on Santa Cruz Island, Galapagos. Prev Vet Med, 123, 128-137.
  • Dorak MT (2007): Real-time PCR. Taylor & Francis. New York.
  • Ek-Kommonen C, Sihvonen L, Pekkanen K, et al (1997): Outbreak of canine distemper in vaccinated dogs in Finland. Vet Rec, 141, 380-383.
  • Elia G, Camero M, Losurdo M, et al (2015): Virological and serological findings in dogs with naturally occurring distemper. J Virol Methods, 213, 127-130.
  • Elia G, Decaro N, Martella V, et al (2006): Detection of canine distemper virus in dogs by real-time RT-PCR. J Virol Methods, 136, 171-176.
  • Esin E (2013): Köpeklerde Canine Distemper Virus enfeksiyonunun araştırılması. Doktora Tezi. Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü, Konya.
  • Fong TT, Lipp EK (2005): Enteric viruses of humans and animals in aquatic environments: health risks, detection, and potential water quality assessment tools. Microbiol Mol Biol Rev, 69, 357-371.
  • Gray LK, Crawford PC, Levy JK, et al (2012): Comparison of two assays for detection of antibodies against canine parvovirus and canine distemper virus in dogs admitted to a Florida animal shelter. J Am Vet Med Assoc, 240, 1084-1087.
  • Greene CE (2013): Infectious diseases of the dog and cat. Elsevier Health Sciences, London.
  • Gustavsson L, Westin J, Andersson LM, et al (2011): Rectal swabs can be used for diagnosis of viral gastroenteritis with a multiple real-time PCR assay. J Clin Virol, 51, 279-282.
  • Headley SA, Sukura A (2009): Naturally occurring systemic canine distemper virus infection in a pup. Braz J Vet Pathol, 2, 95-101.
  • Jóžwik A, Frymus T (2002): Natural distemper in vaccinated and unvaccinated dogs in Warsaw. J Vet Med, 49, 413-414.
  • Kapil S, Neel T (2015): Canine distemper virus antigen detection in external epithelia of recently vaccinated, sick dogs by fluorescence microscopy is a valuable prognostic indicator. J Clin Microbiol, 53, 687-691.
  • Keawcharoen J, Theamboonlers A, Jantaradsamee P, et al (2005): Nucleotide sequence analysis of nucleocapsid protein gene of canine distemper virus isolates in Thailand. Vet Microbiol, 105, 137-142.
  • Lan N, Yamaguchi R, Inomata A, et al (2006): Comparative analyses of canine distemper viral isolates from clinical cases of canine distemper in vaccinated dogs. Vet Microbiol, 115, 32-42.
  • Lednicky JA, Meehan TP, Kinsel MJ, et al (2004): Effective primary isolation of wild-type Canine distemper virus in MDCK, MV1 Lu and Vero cells without nucleotide sequence changes within the entire haemagglutinin protein gene and in subgenomic sections of the fusion and phospho protein genes. J Virol Methods, 118, 147–157.
  • Lorusso A, Savini G (2014): Old diseases for new nightmares: distemper strikes back in Italy. Vet Ital, 50, 151-154.
  • MacLachlan NJ, Dubovi EJ (2016): Paramyxoviridae and pneumoviridae. In: MacLachlan NJ, Dubovi EJ, editors. Fenner’s veterinary virology. 5th ed. London, UK: Academic press, 327-56.
  • Madeley CR, Peiris JSM (2002): Methods in virus diagnosis: immunofluorescence revisited. J Clin Virol, 25, 121–134.
  • Martella V, Cirone, F, Elia G, et al (2006): Heterogeneity within the hemagglutinin genes of canine distemper virus (CDV) strains detected in Italy. Vet Microbiol, 116, 301-309.
  • Martinez-Gutierrez M, Ruiz-Saenz J (2016): Diversity of susceptible hosts in canine distemper virus infection: a systematic review and data synthesis. BMC Vet Res, 12, 78.
  • Nuanualsuwan S, Cliver DO (2002): Pretreatment to avoid positive RT-PCR results with inactivated viruses. J Virol Methods, 104, 217-225.
  • Origgi F, Plattet P, Sattler U, et al (2012): Emergence of canine distemper virus strains with modified molecular signature and enhanced neuronal tropism leading to high mortality in wild carnivores. Vet Pathol, 49, 913-929.
  • Özkul A, Sancak AA, Güngör E, et al (2004): Determination and phylogenetic analysis of canine distemper virus in dogs with nervous symptoms in Turkey. Acta Vet Hung, 52, 125-132.
  • Pawar RM, Raj GD, Gopinath VP, et al (2011): Isolation and molecular characterization of canine distemper virus from India. Trop Anim Health Prod, 43, 1617-1622.
  • Richards GP (1999): Limitations of molecular biological techniques for assessing the virological safety of foods. J Food Prot, 62, 691-697.
  • Roelke-Parker ME, Munson L, Packer C, et al (1996): A canine distemper virus epidemic in Serengeti lions (Panthera leo). Nature, 37, 441–445.
  • Sahna K, Gencay A, Atalay O (2008): Viral aetiology of diarrhoea in puppies from a same shelter in Turkey: presence of mixed infections. Rev Med Vet, 159, 345-346.
  • Saito T, Alfieri A, Wosiacki S, et al (2006): Detection of canine distemper virus by reverse transcriptase-polymerase chain reaction in the urine of dogs with clinical signs of distemper encephalitis. Res Vet Sci, 80, 116-119.
  • Saltık HS, Kale M (2020): Evaluation of infection with N protein-specific Immunoglobulin M and G in naturally occurring distemper in dogs. Vet Med-Czech, 65, 168–173.
  • Schobesberger M, Summerfield A, Doherr MG, et al (2005): Canine distemper virus-induced depletion of uninfected lymphocytes is associated with apoptosis. Vet Immunol Immunopathol, 104, 33-44.
  • Seki F, Ono N, Yamaguchi R (2003): Efficient isolation of wild strains of canine distemper virus in Vero cells expressing canine SLAM (CD150) and their adaptability to marmoset B95a cells. J Virol, 77, 9943-9950.
  • Shin YJ, Cho KO, Cho HS, et al (2004): Comparison of one-step RT-PCR and a nested PCR for the detection of canine distemper virus in clinical samples. Aust Vet J, 82, 83-86.
  • Swati, Deka D, Uppal SK, et al (2015): Isolation and phylogenetic characterization of Canine distemper virus from India. VirusDisease, 26, 133-140.
  • Tan B, Wen YJ, Wang FX, et al (2011): Pathogenesis and phylogenetic analyses of canine distemper virus strain ZJ7 isolate from domestic dogs in China. Virol J, 8, 1.
  • Temilade BE, Solomon OOO, Omotayo OE, et al (2015): Seropositivity of Canine Distemper Virus (CDV) in Dogs Presenting at Abeokuta, Nigeria. Public Health Research, 5, 109-119.
  • Wyllie SE, Kelman M, Ward MP (2016): Epidemiology and clinical presentation of canine distemper disease in dogs and ferrets in Australia, 2006–2014. Aust Vet J, 94, 215-222.

Year 2023, , 49 - 56, 30.12.2022

Hasbi Sait Saltık , Mehmet Kale

https://doi.org/10.33988/auvfd.846475

Abstract

Project Number

0403-DR-16

References

  • Anderson DE, Wang LF (2011): New and emerging paramyxoviruses. 435-459. In: SK Samal (Ed), The Biology of Paramyxoviruses. Caister Academic Press, Maryland, USA
  • Appel MJ, Gillespie JH (1972): Canine distemper virus. 1-96. In: SGC Hallauer and K Meyer (Ed), Virology Monographs. Springer, Vienna.
  • Appel MJG, Summers B (1999): Canine distemper: current status. 68-72. In: L Carmichael (Ed), Recent Advances in Canine Infectious Diseases. International Veterinary Information Service (IVIS).
  • Appel MJG, Summers BA (1995): Pathogenicity of morbilliviruses for terrestrial carnivores. Vet Microbiol, 44, 187-191.
  • Arya M, Shergill IS, Williamson M, et al (2005): Basic principles of real-time quantitative PCR. Expert Rev Mol Diagn, 5, 209-219.
  • Beineke A, Puff C, Seehusen F, et al (2009): Pathogenesis and immunopathology of systemic and nervous canine distemper. Vet Immunol Immunopathol, 127, 1-18.
  • Blancou J (2004): Dog distemper: imported into Europe from South America. Hist Med Vet, 29, 35-41.
  • Blixenkrone-Moeller M, Svansson V, Have P, et al (1993): Studies on manifestations of canine distemper virus infection in an urban dog population. Vet Microbiol, 37, 163-173.
  • Calderon MG, Remorini P, Periolo O, et al (2007): Detection by RT-PCR and genetic characterization of canine distemper virus from vaccinated and non-vaccinated dogs in Argentina. Vet Microbiol, 125, 341-349.
  • Chappuis G (1995): Control of canine distemper. Vet Microbiol, 44, 351-358.
  • Demeter Z, Palade EA, Rusvai M (2009): Canine distemper: Still a major concern in Central Europe. Lucr Stiint Univ Stiint Agric Banat Timis Med Vet, 42, 136-150.
  • Di Sabatino D, Savini G, Lorusso A (2015): Canine distemper and endangered wildlife: Is it time for mandatory vaccination of dogs. Vaccine, 33, 6519.
  • Diaz NM, Mendez GS, Grijalva CJ, et al (2016): Dog overpopulation and burden of exposure to canine distemper virus and other pathogens on Santa Cruz Island, Galapagos. Prev Vet Med, 123, 128-137.
  • Dorak MT (2007): Real-time PCR. Taylor & Francis. New York.
  • Ek-Kommonen C, Sihvonen L, Pekkanen K, et al (1997): Outbreak of canine distemper in vaccinated dogs in Finland. Vet Rec, 141, 380-383.
  • Elia G, Camero M, Losurdo M, et al (2015): Virological and serological findings in dogs with naturally occurring distemper. J Virol Methods, 213, 127-130.
  • Elia G, Decaro N, Martella V, et al (2006): Detection of canine distemper virus in dogs by real-time RT-PCR. J Virol Methods, 136, 171-176.
  • Esin E (2013): Köpeklerde Canine Distemper Virus enfeksiyonunun araştırılması. Doktora Tezi. Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü, Konya.
  • Fong TT, Lipp EK (2005): Enteric viruses of humans and animals in aquatic environments: health risks, detection, and potential water quality assessment tools. Microbiol Mol Biol Rev, 69, 357-371.
  • Gray LK, Crawford PC, Levy JK, et al (2012): Comparison of two assays for detection of antibodies against canine parvovirus and canine distemper virus in dogs admitted to a Florida animal shelter. J Am Vet Med Assoc, 240, 1084-1087.
  • Greene CE (2013): Infectious diseases of the dog and cat. Elsevier Health Sciences, London.
  • Gustavsson L, Westin J, Andersson LM, et al (2011): Rectal swabs can be used for diagnosis of viral gastroenteritis with a multiple real-time PCR assay. J Clin Virol, 51, 279-282.
  • Headley SA, Sukura A (2009): Naturally occurring systemic canine distemper virus infection in a pup. Braz J Vet Pathol, 2, 95-101.
  • Jóžwik A, Frymus T (2002): Natural distemper in vaccinated and unvaccinated dogs in Warsaw. J Vet Med, 49, 413-414.
  • Kapil S, Neel T (2015): Canine distemper virus antigen detection in external epithelia of recently vaccinated, sick dogs by fluorescence microscopy is a valuable prognostic indicator. J Clin Microbiol, 53, 687-691.
  • Keawcharoen J, Theamboonlers A, Jantaradsamee P, et al (2005): Nucleotide sequence analysis of nucleocapsid protein gene of canine distemper virus isolates in Thailand. Vet Microbiol, 105, 137-142.
  • Lan N, Yamaguchi R, Inomata A, et al (2006): Comparative analyses of canine distemper viral isolates from clinical cases of canine distemper in vaccinated dogs. Vet Microbiol, 115, 32-42.
  • Lednicky JA, Meehan TP, Kinsel MJ, et al (2004): Effective primary isolation of wild-type Canine distemper virus in MDCK, MV1 Lu and Vero cells without nucleotide sequence changes within the entire haemagglutinin protein gene and in subgenomic sections of the fusion and phospho protein genes. J Virol Methods, 118, 147–157.
  • Lorusso A, Savini G (2014): Old diseases for new nightmares: distemper strikes back in Italy. Vet Ital, 50, 151-154.
  • MacLachlan NJ, Dubovi EJ (2016): Paramyxoviridae and pneumoviridae. In: MacLachlan NJ, Dubovi EJ, editors. Fenner’s veterinary virology. 5th ed. London, UK: Academic press, 327-56.
  • Madeley CR, Peiris JSM (2002): Methods in virus diagnosis: immunofluorescence revisited. J Clin Virol, 25, 121–134.
  • Martella V, Cirone, F, Elia G, et al (2006): Heterogeneity within the hemagglutinin genes of canine distemper virus (CDV) strains detected in Italy. Vet Microbiol, 116, 301-309.
  • Martinez-Gutierrez M, Ruiz-Saenz J (2016): Diversity of susceptible hosts in canine distemper virus infection: a systematic review and data synthesis. BMC Vet Res, 12, 78.
  • Nuanualsuwan S, Cliver DO (2002): Pretreatment to avoid positive RT-PCR results with inactivated viruses. J Virol Methods, 104, 217-225.
  • Origgi F, Plattet P, Sattler U, et al (2012): Emergence of canine distemper virus strains with modified molecular signature and enhanced neuronal tropism leading to high mortality in wild carnivores. Vet Pathol, 49, 913-929.
  • Özkul A, Sancak AA, Güngör E, et al (2004): Determination and phylogenetic analysis of canine distemper virus in dogs with nervous symptoms in Turkey. Acta Vet Hung, 52, 125-132.
  • Pawar RM, Raj GD, Gopinath VP, et al (2011): Isolation and molecular characterization of canine distemper virus from India. Trop Anim Health Prod, 43, 1617-1622.
  • Richards GP (1999): Limitations of molecular biological techniques for assessing the virological safety of foods. J Food Prot, 62, 691-697.
  • Roelke-Parker ME, Munson L, Packer C, et al (1996): A canine distemper virus epidemic in Serengeti lions (Panthera leo). Nature, 37, 441–445.
  • Sahna K, Gencay A, Atalay O (2008): Viral aetiology of diarrhoea in puppies from a same shelter in Turkey: presence of mixed infections. Rev Med Vet, 159, 345-346.
  • Saito T, Alfieri A, Wosiacki S, et al (2006): Detection of canine distemper virus by reverse transcriptase-polymerase chain reaction in the urine of dogs with clinical signs of distemper encephalitis. Res Vet Sci, 80, 116-119.
  • Saltık HS, Kale M (2020): Evaluation of infection with N protein-specific Immunoglobulin M and G in naturally occurring distemper in dogs. Vet Med-Czech, 65, 168–173.
  • Schobesberger M, Summerfield A, Doherr MG, et al (2005): Canine distemper virus-induced depletion of uninfected lymphocytes is associated with apoptosis. Vet Immunol Immunopathol, 104, 33-44.
  • Seki F, Ono N, Yamaguchi R (2003): Efficient isolation of wild strains of canine distemper virus in Vero cells expressing canine SLAM (CD150) and their adaptability to marmoset B95a cells. J Virol, 77, 9943-9950.
  • Shin YJ, Cho KO, Cho HS, et al (2004): Comparison of one-step RT-PCR and a nested PCR for the detection of canine distemper virus in clinical samples. Aust Vet J, 82, 83-86.
  • Swati, Deka D, Uppal SK, et al (2015): Isolation and phylogenetic characterization of Canine distemper virus from India. VirusDisease, 26, 133-140.
  • Tan B, Wen YJ, Wang FX, et al (2011): Pathogenesis and phylogenetic analyses of canine distemper virus strain ZJ7 isolate from domestic dogs in China. Virol J, 8, 1.
  • Temilade BE, Solomon OOO, Omotayo OE, et al (2015): Seropositivity of Canine Distemper Virus (CDV) in Dogs Presenting at Abeokuta, Nigeria. Public Health Research, 5, 109-119.
  • Wyllie SE, Kelman M, Ward MP (2016): Epidemiology and clinical presentation of canine distemper disease in dogs and ferrets in Australia, 2006–2014. Aust Vet J, 94, 215-222.
There are 49 citations in total.

Details

Primary Language English
Subjects Veterinary Surgery
Journal Section Research Article
Authors

Hasbi Sait Saltık 0000-0002-3283-7062

Mehmet Kale 0000-0003-4156-1077

Project Number 0403-DR-16
Publication Date December 30, 2022
Published in Issue Year 2023

Cite

APA Saltık, H. S., & Kale, M. (2022). Rapid molecular detection and isolation of canine distemper virus in naturally infected dogs. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 70(1), 49-56. https://doi.org/10.33988/auvfd.846475
AMA Saltık HS, Kale M. Rapid molecular detection and isolation of canine distemper virus in naturally infected dogs. Ankara Univ Vet Fak Derg. December 2022;70(1):49-56. doi:10.33988/auvfd.846475
Chicago Saltık, Hasbi Sait, and Mehmet Kale. “Rapid Molecular Detection and Isolation of Canine Distemper Virus in Naturally Infected Dogs”. Ankara Üniversitesi Veteriner Fakültesi Dergisi 70, no. 1 (December 2022): 49-56. https://doi.org/10.33988/auvfd.846475.
EndNote Saltık HS, Kale M (December 1, 2022) Rapid molecular detection and isolation of canine distemper virus in naturally infected dogs. Ankara Üniversitesi Veteriner Fakültesi Dergisi 70 1 49–56.
IEEE H. S. Saltık and M. Kale, “Rapid molecular detection and isolation of canine distemper virus in naturally infected dogs”, Ankara Univ Vet Fak Derg, vol. 70, no. 1, pp. 49–56, 2022, doi: 10.33988/auvfd.846475.
ISNAD Saltık, Hasbi Sait - Kale, Mehmet. “Rapid Molecular Detection and Isolation of Canine Distemper Virus in Naturally Infected Dogs”. Ankara Üniversitesi Veteriner Fakültesi Dergisi 70/1 (December 2022), 49-56. https://doi.org/10.33988/auvfd.846475.
JAMA Saltık HS, Kale M. Rapid molecular detection and isolation of canine distemper virus in naturally infected dogs. Ankara Univ Vet Fak Derg. 2022;70:49–56.
MLA Saltık, Hasbi Sait and Mehmet Kale. “Rapid Molecular Detection and Isolation of Canine Distemper Virus in Naturally Infected Dogs”. Ankara Üniversitesi Veteriner Fakültesi Dergisi, vol. 70, no. 1, 2022, pp. 49-56, doi:10.33988/auvfd.846475.
Vancouver Saltık HS, Kale M. Rapid molecular detection and isolation of canine distemper virus in naturally infected dogs. Ankara Univ Vet Fak Derg. 2022;70(1):49-56.