Paper Details

Detection of newcastle disease virus from wild quail (Coturnix coturnix) by reverse transcription polymerase chain reaction in Isfahan and Loretan Provinces of Iran

Forutan Salehinezhad, Amir Shafiei, Hossein Faraji Khoo, Sheida Zarasvand Bagheri, NastaranAbdi Dezfuli

DOI: https://dx.doi.org/10.12692/ijb/4.2.141-147

Int. J. Biosci. 4(2), 141-147. January, 2014. (PDF)

Abstract:

The present research work was carried out to isolate and identify Newcastle disease virus (NDV) by using haemagglutination inhibition (HI) test and reverse transcription-polymerase chain reaction (RT-PCR) assay. A total of 500 clinical and post-mortem (brain, lung, colon and spleen) samples were collected from of two populations of Coturnix Coturnix of Newcastle disease (ND) in 2013, one in Lorestan province and other one in Isfahan province . All the samples were inoculated onto 10-day-old embryonated through allantoic sac route and in the chicken embryo fibroblasts (CEFs) cell culture. The allantoic fluid (AF) of the dead embryos and the infected culture fluid (ICF) of the CEF were harvested at 48 and 96 hours of post-infection,respectively. The HI and RT-PCR were employed to detect NDV in tissue homogenates of all the clinical and post-mortem samples as well as laboratory samples (AF and ICF). In CEF cell culture system, the rate of virus isolation from all the aforesaid samples was found 100% with the exception of serum samples. The isolation rate of NDV was higher in CEF culture system (93.8%) compared to that of avian embryos (80%). The anti-NDV hyperimmune serum revealed complete inhibition of the 4 haemagglutination unit of each isolate of viruses isolated from broiler and layer chickens present in the laboratory samples (AF and ICF). The NDV specific primers used in the direct RT-PCR for genome detection of NDV showed equal sensitivity and specificity with the RNA extracted from the clinical, post-mortem and laboratory samples (AF and ICF) as with the genomic RNA of reference NDV. Higher rate of detection of NDV was recorded with RT-PCR assay than HI test. Therefore, the molecular method (RT-PCR) can be introduced for rapid and confirmatory detection of NDV from different forms of Isfahan and Lorestan province but there is no report on Newcastle disease Coturnix Coturnix in Iran.