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In the present study, fowl pox was diagnosed in 129 backyard chicken based on gross and histopathological lesions. Fowl pox is manifested by pock nodular lesions on featherless parts of body (cutaneous form) and purulent, cheesy patches in upper respiratory tract (diphtheritic form). Tissue samples revealed Bollinger bodies in epithelial cells on histopathology. Among these, extraction of DNA was carried out from cutaneous and diphtheretic lesions from 29 fowl pox affected chicken. Molecular diagnosis of fowl pox was performed by amplification of P4b gene of fowl pox virus using specific primers and obtained a desired amplicon of 578 bp. Nucleotide sequencing and phylogenetic analysis of four cutaneous fowl pox samples revealed close relation with the FPV isolates from Singapore, North America and USA (KY464130, MH175285 and KC017961 with an identity of 97.28%, 97.21% and 97.16% respectively) when compared to FPVs isolated from Portugal, Brazil, Jammu and India (KM974727, KX863706, MF496042 and HM481406 respectively with an identity of 96.66% to 96.86%). [ABSTRACT FROM AUTHOR]

Fowlpox is a re-emerging and economically significant viral disease in poultry. Fowlpox is a complex virus contains a double-strand DNA genome of approximately 300 kb and largest among all pox viruses. In the present study, a clinical case of fowlpox virus (FPV) infection was investigated and a Real-Time PCR (RT-PCR) assay was developed for confirmatory diagnosis of fowl pox and quantification of viral load. Clinical sample was collected from infected bird reared in the organized poultry farm management system and the viral infection was confirmed using conventional PCR amplification of the P4b gene of fowl pox virus. Following that, a single-step SYBR green based Real-Time PCR assay was developed by targeting the same gene of fowlpox virus. The fowl pox viral load in the infected bird was found to be 7.33 ± 0.044 (Log Mean ± SD). The lower detection limit achieved was 134 copies per microliter (µL) with a corresponding threshold cycle (Ct) value of 32.59. Thus, the single-step SYBR green based RT-PCR assay targeting the P4b gene of fowl pox virus reported in this study was highly sensitive, specific and reproducible for the detection and quantitation of fowl pox virus nucleic acids. [ABSTRACT FROM AUTHOR]