Abstract
Many viruses have been associated with runting and stunting syndrome (RSS). These viral infections mainly affect young chickens, causing apathy, depression, ruffled feathers, cloacal pasting, and diarrhea. Chicken Parvovirus (ChPV) is such an infection and has been detected in chickens showing signs of enteric diseases worldwide. Therefore, the present study aims to develop a sensitive real-time fast-qPCR assay based on SYBR® Green for detection and quantification of ChPV. A 561-bp non-structural (NS) gene was amplified and cloned, and a pair of primers was designed based on conserved nucleotide sequences on the NS gene of ChPV, the intercalating DNA reagent SYBR® Green was employed, and the Fast mode of a thermocycler was used. The assay detects 109 to 101 copies of the genome (CG). The limit of detection (LoD) was estimated to five CG, and the limit of quantification (LoQ) was estimated at ten CG. The standard curve efficiency was 101.94%, and the melting curve showed a unique clean peak and a melting temperature of 79.3 °C. The assay was specific to amplify the ChPV NS gene, and no amplification was shown from other viral genomes or in the negative controls. A total of 141 samples were tested using the assay, of which 139 samples were found positive. The highest CG value of ChPV was 5.7 × 106 CG/uL of DNA without apparent clinical signs of enteric disturbance, and 4.6 × 106 CG/uL DNA were detected in chickens with RSS.
Highlights
Runting-stunting syndrome (RSS) is a pathogen that afflicts young, one-week-old chicks, causing poor feed conversion, stunting, dwarfism, culling and mortality [1,2,3]
We report the development of a highly sensitive real-time fast-qPCR assay based on SYBR® Green for detection and quantification of Chicken Parvovirus (ChPV), which is faster than other current assays due its combination of cycle steps, faster changes in temperature, faster enzymes, and is so sensitive that very low concentrations of the virus present in the enteric contents of animals with or without clinical signs of enteric disease can be detected
Gene obtained in the present study showed a high similarity with other ChPV sequences from other parts of the world, clustering with ChPV, and one sequence clustered with the Turkey Parvovirus (TuPV), but this result does not indicate that this sequence belongs to TuPV [28]
Summary
Runting-stunting syndrome (RSS) is a pathogen that afflicts young, one-week-old chicks, causing poor feed conversion, stunting, dwarfism, culling and mortality [1,2,3]. The chicks present with depression, apathy, ruffled feathers, cloacal pasting, and the most characteristic signs, diarrhea and unsteadiness [3,4]. The etiological agents related to RSS remain unknown; many viruses have been detected and implicated as causative agents of the disease, including astroviruses, coronaviruses, rotaviruses, reovirus, parvovirus, and others [5,6,7,8,9,10]. Parvovirus is a viral agent that affects many animals, including mammals such as cows, dogs, cats, rats, and bats [11].
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
