Journal of Food Quality and Hazards Control
مجله کیفیت و کنترل مخاطرات مواد غذایی
J. Food Qual. Hazards Control
Medical Sciences
http://jfqhc.ssu.ac.ir
1
admin
2345-685X
2345-6825
8
10.29252/jfqhc
14
8888
13
en
jalali
1395
3
1
gregorian
2016
6
1
3
2
online
1
fulltext
en
Development of a Differential PCR Assay for Detection of Brucella abortus and Brucella melitensis: an Analytical Approach for Monitoring of Brucella spp. in Foods of Animal Origin
تخصصي
Special
Original article
Original article
<p style="text-align: justify;"><span style="font-family:times new roman;"><strong>Background: </strong>Classical bacteriological detection of <em>Brucella</em> species from food, and environment is routinely carried out based on morphological and biochemical characteristics. However, for increasing specificity and sensitivity of species identification methods, development of a molecular assay is necessary that was main aim of this study.<br>
<strong>Methods: </strong>Panel of some reference strains belonging to the phylum <em>Proteobacteria </em>were specifically used in this study. Additionally, the panel was enriched with 20 <em>Brucella</em> field strains isolated from 13 cattle and 7 sheep (West Kazakhstan region), and six strains from three cattle and three sheep (Almaty region). Bacterial identification before designing was carried out based on <em>16S rRNA </em>sequencing with universal primers. Primer design was implemented using the Primer3 program. Finally, specificity and sensitivity of the PCR assay for <em>Brucella</em> identification were evaluated.</span></p>
<p style="text-align: justify;"><span style="font-family:times new roman;"><strong>Results: </strong>The sensitivity of the developed conventional PCR assays was assessed with the range of 2×10<sup>5</sup> to 12 genomic copies isolated from <em>B. abortus </em>100 and <em>B. melitensis</em> H-12 reference strains. The sensitivity of the developed assays using Ba and Ba-r, Bm and Bm-r primers was determined to be 1.6×10<sup>3 </sup>genomic copies.</span></p>
<p style="text-align: justify;"><span style="font-family:times new roman;"><strong>Conclusion: </strong>Quick detection and species identification of <em>Brucella</em> strains circulating in Kazakhstan would help local authorities in decision-making and implementation of the most effective strategies for control of these bacteria. Our PCR-based assay was the first step towards developing a novel kit with final aim of standardizing molecular identification of <em>B. abortus</em> and <em>B. melitensis</em> in foods of animal origin in Kazakhstan and other central Asia countries.</span></p>
Polymerase Chain Reaction, Brucella, Food Safety
53
59
http://jfqhc.ssu.ac.ir/browse.php?a_code=A-10-235-13&slc_lang=en&sid=1
А.
Daugaliyeva
aida1979@bk.ru