Title: PCR BASED P32 GENE IS A CONFIRMATORY DIAGNOSTIC TECHNIQUE FOR SHEEP AND GOAT POX VIRUS

Authors: Amol S Kinhekar, VC Ingle and PA Tembhurne

Source: Ruminant Science (2017)-6(1):75-78.

Cite this reference as: Kinhekar Amol S, Ingle VC and Tembhurne PA (2017). PCR based P32 gene is a confirmatory diagnostic technique for sheep and goat pox virus. Ruminant Science 6(1):75-78.

Abstract

Efforts are being held for identifying suitable diagnostic tool in respect to contagious pox virus affecting small ruminants. In this respect, this study was carried out to diagnose and confirm suspected outbreak of Sheep and Goat pox based on P32 gene, the most conserved region of capripox genome. Scab samples were collected and preserved from suspected clinical outbreaks among small ruminants in regions of Maharashtra, INDIA were cultivated on Vero CCL-81cell culture. Observation during third passage revealed cytopathic effect (CPE) viz, cell ballooning and detachment. DNA isolation was carried out from these cell lines for diagnosis of infection through polymerase chain reaction depending on the size of the amplicon corresponding to the target gene (1000 bp). The amplified P32 gene from designated primer confirms the presence of sheep and goat pox virus in these biopsy materials. The current study had confirmed that P32 gene based PCR assay is a rapid and accurate diagnostic tool for detection of sheep and goat pox virus from clinical samples.

References

Amal MA, Raof MA, Shahein and Hoda AEM (2008). Electrophoretic characterization of sheep pox virus, goat pox virus and lumpy skin disease virus. Egyptain Journal of Comparative Pathology and Clinical Pathology 21(1):15-30.

Babiuk S, Parkyn G, Copps J, Larence JE, Sabara MI, Bowden TR, Boyle DB and Kitching RP (2007). Evaluation of an ovine testis cell line (OA3.Ts) for propagation of capripoxvirus isolates and development of an immunostaining technique for viral plaque visualization. Journal of Veterinary Diagnostic Investigation 19:486-491.

Batra K, Kumar A, Kumar V, Nanda T, Maan NS and Maan S (2015). Development and evaluation of loop-mediated isothermal amplification assay for rapid detection of capripoxvirus. Veterinary World 8(11):1286-1292.

Beard PM, Sugar S, Bazarragchaa E, Gerelmaa U, Tserendorj S, Tuppurainen E and Sodnomdarjaa R (2010). A description of two outbreaks of capripoxvirus disease in Mongolia. Veterinary Microbiology 142(3-4):427-431.

Bhanuprakash V, Venkatesan G, Balamurugan V, Hosamani M, Yogisharadhya R, Chauhan RS, Pande A, Mondal B and Singh RK (2010). Pox outbreaks in sheep and goats at Makhdoom (Uttar Pradesh), India: Evidence of sheeppox virus infection in goats. Transboundary and Emerging Diseases 57(5):375-82.

Buller RM, Arif BM, Black DN, Dumbell KR, Esposito JJ, Lefkowitz EJ, McFadden G, Moss B, Mercer AA, Moyer RW, Skinner MA and Tripathy DN (2005). family poxviridae. virus taxonomy: Classification and nomenclature of viruses. Eighth Report of the International Committee on Taxonomy of Viruses 117-133.

Chand P, Kitching RP and Black DN (1994). Western blot analysis of virus-specific antibody responses for capripox and contagious pustular dermatitis viral infections in sheep. Epidemiology and Infection 113:377-385.

Chandranaik BM, Manjunath L, Venkatesha MD, Muniyellappa HK, Giridhar P and Renukaprasad C (2014). Isolation of goatpox virus and application of polymerase chain reaction for diagnosis of goat pox from scab materials. Indian Veterinary Journal 91(2):90-91.

Das A, Babiuk S and McIntosh MT (2012). development of a loop-mediated isothermal amplification assay for rapid detection of capripox viruses. Journal of Clinical Microbiology 50(5):1613-1620.

Diallo A and Viljoen GJ (2007). Genus Capripox. Birkhäuser Advances in Infectious Diseases 167-181.

Fulzele S, Singh RK and Hosamani M (2006). Evolution of duplex PCR and PCR-RFLP for diagnosis of sheep pox and goat pox. International Journal of Tropical Medicine 1(2):66-70.

Garner MG, Sawarkar SD, Brett EK, Edwards JR, Kulkarni VB, Boyle DB and Singh SN (2000). The extent and impact of sheep pox and goat pox in the state of Maharashtra, India. Tropical Animal Health Production 32:205-223.

Heine HG, Stevens MP, Foord AJ and Boyle DB (1999). A capripoxvirus detection PCR and antibody ELISA based on the major antigen P32, the homolog of the vaccinia virus H3L gene. Journal of Immunological Methods 227:187-196.

Hosamani M, Mondal B, Tembhurne PA, Bandopadhyay SK, Singh RK and Rasool TJ (2004). Differentiation of sheep pox and goat poxviruses by sequence analysis and PCR-RFLP of P32 gene.  Virus Genes 29(1):73-80.

Ireland DC and Binepal YS (1998). Improved detection of capripoxvirus in biopsy samples by PCR. Journal of Virological Methods 74:1-7.

Kinhekar AS, Raja P, Tembhurne PA, Ingle VC, Manesh Kumar P and Kalorey DR (2012). P32 gene based molecular characterization of Sheep pox and Goat pox viruses. International symposium on One health: Way forwards to challenges in food safety and zoonoses in 21st century and XI annual conference of Indian veterinary public health specialist (IAVPHS), organized by GADVASU, Ludhiana, Punjab, 13-14, December.

Mangana-Vougiouka O, Markoulatos P, Koptopoulos G, Nomikou K, Bakandritsos N and Papadopoulos O (1999). Sheep poxvirus identification by PCR in cell cultures. Journal of Virological Methods 77:75-79.

Mirzaie K, Barani SM and Bokaie S (2015). A review of sheep pox and goat pox: Perspective of their control and eradication in Iran. Journal of Advancement in Veterinary and Animal Research 2(4):373-381.

Murray L, Edwards L, Tuppurainen ESM, Bachanek-Bankowska K, Oura CAL and Mioulet V (2013). Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay. BMC Veterinary Research 9:90.

Roy P, Purushothaman V, Sreekumar C, Tamizharasan S and Chandramohan A (2008). Sheep pox disease outbreaks in Madras Red and Mechery breeds of indigenous sheep in Tamilnadu, India. Research in Veterinary Science 85:617-621.

Sambrook J, Fritsch EF and Maniatis T (1989). Molecular Cloning: A Laboratory Manual, Vol. 3. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

Sharma R, Patil RD, Parimoo HA , Thakur D and Katoch VC (2013). Clinico-pathology of sheep pox disease in Himachal Pradesh, India. Ruminant Science 2(2):127-130.

Tian H, Chen Y, Wu J, Shang Y and Liu X (2010). Serodiagnosis of sheeppox and goatpox using an indirect ELISA based on synthetic peptide targeting for the major antigen P32. Virology Journal 7:245.

Varshovi HR, Keyvanfar H, Aghaiypour K, Pourbakhsh SA, Shooshtari AH and Aghaebrahimian M (2009). Capripoxvirus identification by PCR based on P32 gene. Archives of Razi Institute 64(1):19-25.

Yune N and Abdela N (2017). Epidemiology and economic importance of sheep and goat pox: A review on past and current aspects. Journal of Veterinary Science and Technology 8:2.

Zhou T, Chen HG, He X, Yongxiang FY, Wang Guan Q, Zeng S, Cui Q and Jing Z (2012). Phylogenetic analysis of Chinese sheeppox and goatpox virus isolates. Virology Journal 9:25.