Detection and characterization of Newcastle disease virus in formalin-fixed, paraffin-embedded tissues from commercial broilers in Egypt

Mostafa Y. Abdel-Glil, Sunil K. Mor, Tamer A. Sharafeldin, Robert E. Porter, Sagar M. Goyal

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Newcastle disease (ND) is highly contagious and causes severe economic losses to the poultry industry due to high morbidity and mortality. In this report, we describe the detection of Newcastle disease virus (NDV) in formalin-fixed tissues from an outbreak of ND on broiler farms in Egypt. The affected birds experienced respiratory and/or nervous signs and a 75% mortality rate. Tissue samples were collected and placed in 10% neutral buffered formalin followed by embedding in paraffin. RNA was extracted from 80-m formalin-fixed paraffin-embedded tissue blocks and recovered in 60 l of elution buffer. All samples were negative for influenza virus by real-time reverse-transcription (RT)-PCR but positive for NDV. These flocks were known to have been vaccinated with a live NDV vaccine (LaSota strain). The nucleic acid sequences of the virus detected in this study were similar to those of a velogenic virus at its cleavage site 111GRRQKR*F117 and clustered with class II genogroup VII lineage of NDV, with a nucleotide sequence identity of 94%-99%. Although extraction and amplification of NDV from paraffin-embedded tissues from experimentally infected birds has been reported previously, this study reports on the use of RT-PCR on formalin-fixed tissues from actual field samples.

Original languageEnglish (US)
Pages (from-to)118-123
Number of pages6
JournalAvian diseases
Volume58
Issue number1
DOIs
StatePublished - Mar 2014

Keywords

  • Egypt
  • Newcastle disease virus
  • RT-PCR
  • formalin-fixed tissues
  • genotype VII

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