Keywords : Polymerase chain reaction


Isolation and identification of Circovirus in pigeon

Safwan Yousif Al-Baroodi; Mozahim yasen Al-Attar

Iraqi Journal of Veterinary Sciences, In Press
DOI: 10.33899/ijvs.2020.126706.1364

The purpose of this study is first trial to detect of pigeon circovirus, so 1sr group include 100 cloacal swabs were collected 55 healthy and 45 ill pigeons, 36 yearlings and 64 adults, the 2nd group included organs was liver, spleen, bursa of Fabricius from 41 young pigeons 10-30 days old and bursa of Fabricius, liver, spleen from 28 dead in shell pigeon embryo in the 3rd group. DNA extracted from this samples and detection of virus DNA was attempt using polymerase chain reaction, after DNA amplification, the final products of the amplicon with 331 bp was cleared by using electrophoresis using agarose gel at concentration 2%. Results of viral DNA amplification were positive, which revealed as band in 331 bp the results showed that ill yearling pigeons recording high infectivity rate (66.7%)compare with healthy yearling pigeons and adult once, the bursa of Fabricius samples of dead yearling pigeons recorded high prevalence (36.58%) when compare with liver and spleen samples, DNA of pigeon circovirus high detected (60.71%) in bursa of Fabricius of dead in shell pigeon embryo. inconclusion pigeon circovirus affected the racing pigeon in Mosul, Iraq.

Detection of Shigella in raw bovine milk by polymerase chain reaction

Noor Soulieman; Aemaan Al-Mariri; Faizah Al-Atrash

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 9-16
DOI: 10.33899/ijvs.2019.125758.1146

Shigella is an intracellular bacterium can infect both human and animal. Its species especially Shigella dysenteriae cause shigellosis worldwide, with 165 million cases of severe bloody diarrhea and mucoid feces. The aim of this study was to find a rapid, sensitive and specific method for screening Shigella in raw bovine contaminated milk. For this goal, 70 samples of milk collected in sterile containers for isolating of Shigella and culturing it on selective media to identify and characterize its morphology, biochemical and molecular characteristics. This study was compared between three different DNA extraction techniques for polymerase chain reaction (direct DNA extraction using a kit, alkaline DNA extraction, and filtrated milk). Our results showed that PCR was able to detect Shigella in 15 out of 15 cases after the milk samples filtered. In other words, the filter technique can be used to detect Shigella in contaminated milk.

Different vectors used to transform and clone of nonstructural NS1 gene of Influenza B in Escherichia coli

Ali Dawood

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 329-333
DOI: 10.33899/ijvs.2019.162964

Flu is a highly contagious and common illness caused by influenza A, B, and C viruses. The aim of the present study was to investigate the transformation and cloning of NS1B gene with pET-32a, pET-32b and pQE-81L in Escherichia coli BL21(DE3) and DH5α. pUC57-NS1B synthetic gene was transform and clone in Escherichia coli BL21(DE3). Isolation, single digestion and ligation with pET-32b using HindIII restriction enzyme. Amplification of recombinant DNA was done with conventional PCR after transformation. Screening with IPTG of colonies. Gel electrophoresis was done for each step of cloning after isolation. Isolation, double digestion and ligation with pET-32a and pQE-81L using SacI, PstI and HindIII respectively.Recombinant DNA was attempted to be transformed into E. coli strains BL21 (DE3) and DH5α. pUC57 plasmid carrying NS1B gene was successful transformed and isolated from E. coli BL21 (DE3). Designed primers used for PCR of NS1B showed successful amplification. First screening of pET-32b-NS1B colonies using white/blue method, cloning NS1B into pET-32b using single restriction digestion with HindIII, pET-32a using double restriction digestion with SacI and HindIII and pQE-81L using double restriction digestion with PstI and HindIII gave unexpected result. This result may relate to re-ligation of digested vector for single digestion and uncompleted digestion for vectors of double restriction digestion. Current study has suggested that recombinant NS1B gene can be cloned using single digestion with other expression vectors.